US20130252950A1

US20130252950A1 - Targeting mtor substrates in treating proliferative diseases

Info

Publication number: US20130252950A1
Application number: US13/825,723
Authority: US
Inventors: John Blenis; Steven P. Gygi; Yonghao Yu
Original assignee: Harvard College
Current assignee: Harvard College
Priority date: 2010-09-23
Filing date: 2011-09-23
Publication date: 2013-09-26
Also published as: WO2012040602A3; WO2012040602A2

Abstract

Provided are over 300 mTOR kinase targets identified by a comprehensive phosphoproteomics assay. Methods of targeting mTOR kinase targets, methods to determine the level of mTOR activity by measuring the level of phosphorylation of an mTOR targeted phosphorylation site, methods for distinguishing different classes of mTOR activity in a cell based on phosphoproteomic analysis of mTOR-targeted proteins are also provided. Also provided is the classification of a hyperproliferative disease based on phosphoproteomic analysis of mTOR-targeted proteins, as well as the personalization of therapeutic methods for the treatment of hyperproliferative disease based on phosphoproteomics. Also provided are therapeutic methods including administering to a subject an mTOR inhibitor, an mTOR inhibitor and an additional kinase inhibitor, or a dual inhibitor of mTOR and an additional kinase based on the phosphorylation levels of mTOR targets determined in the subject. Some aspects of this invention relate to the discovery that GrblO is an mTOR-targeted tumor suppressor gene.

Description

RELATED APPLICATIONS

This application claims priority under 35 U.S.C. §119(e) to U.S. provisional patent application, U.S. Ser. No. 61/403,932, filed Sep. 23, 2010, which is incorporated herein by reference.

GOVERNMENT SUPPORT

This invention was made with U.S. Government support under grant numbers GM051405 and HG3456 awarded by the National Institutes of Health. The U.S. Government has certain rights in the invention.

BACKGROUND

mTOR is an evolutionarily conserved ser/thr protein kinase that controls many critical cellular processes including growth, protein translation, metabolic flux, and cell survival. mTOR functions as the core catalytic component of two structurally and functionally distinct signaling complexes. mTOR complex 1 (mTORC1) regulates cell growth and is responsible for the well-characterized role of mTOR in controlling protein translation whereas mTOR complex 2 (mTORC2) regulates cell survival and the actin cytoskeleton (1-3). The mechanisms responsible for modulating mTORC1 and mTORC2 activity in response to upstream inputs such as growth factors, energetic status, and amino acid levels have been well studied (1). In contrast, relatively few direct substrates of mTOR have been identified and in many cases the mechanisms underlying mTOR's ability to regulate important aspects of cell biology are not known.
Misregulated mTOR activity is a common feature of most cancers (2). Despite great interest, clinical trials for evaluating the selective mTORC1 inhibitor rapamycin as an anti-cancer agent have met with limited success (3). Rapamycin resistance has emerged as a major challenge to its clinical use (4) and is caused in part by feedback loops that activate the PI3K and MAPK signaling pathways in rapamycin-treated cells through poorly understood mechanisms (5, 6).

SUMMARY OF THE INVENTION

Functional characterization of the mTOR signaling pathways has been hampered by the paucity of substrates that have been identified to date. Identifying novel substrates of mTORC1 and mTORC2 is important for making progress toward our general understanding how mTOR signals to downstream effectors and to specifically define components of the feedback loops involved in rapamycin resistance. The best-characterized mTORC1 substrates include p70S6K and 4EBP, whereas mTORC2 phosphorylates several members of the AGC kinase family, including Akt, SGK, and PKC.
The present invention is based in part on large-scale quantitative phospho-proteomics experiments that were performed to define the entire signaling networks downstream of both mTORC1 and mTORC2. Novel mTORC1 substrates identified herein include, but are not limited to Grb10, FOXK1, ZEB2, NDRG3, LARP1, SRPK2, CDK12, MIB1, and IBTK.
Extensive characterization of a novel mTORC1 substrate, the growth factor receptor-bound protein 10 (Grb10), shows that mTORC1-mediated phosphorylation stabilizes Grb10, leading to feedback inhibition of the PI3K and MAPK pathways. In addition, Grb10 expression was shown to be frequently downregulated in a variety of cancers. Interestingly, loss of Grb10 and the well-established tumor suppressor PTEN are mutually exclusive events. Grb10 has been found to be both a novel mTORC1 substrate and a tumor suppressor with relevance across a broad spectrum of cancer subtypes.
In some aspects, the invention provides methods for determining mTOR kinase activity in a cell, the method comprising obtaining a cancer cell from a subject diagnosed to have a cancer, determining the level of Grb10 phosphorylation in the cell, and comparing the level of Grb10 phosphorylation to a reference level, wherein if the level of Grb10 phosphorylation in the cell is higher than the reference level, then the cell is determined to exhibit an elevated level of mTOR kinase activity. In some embodiments, the cell is a normal or healthy cell. In some embodiments, the cell is a cell obtained from a subject not diagnosed with a neoplastic disease. In some embodiments, the cell is a cell obtained from a subject not diagnosed with a cancer. In some embodiments, the cell is a neoplastic cell. In some embodiments, the cell is a cancer cell. In some embodiments, the cell is a cell obtained from a subject diagnosed with a neoplastic disease, for example, with a cancer.
In some aspects, the invention provides methods for selecting a treatment of a neoplastic disease, for example, of a cancer, in a subject, the method comprising obtaining a cell from a subject diagnosed to have a neoplastic disease, for example, a cancer, exhibiting an elevated level of mTOR activity; determining the level of Grb10 expression in the cell; and comparing the level of Grb10 expression to a reference level, wherein if the level of Grb10 expression in the cell is higher than the reference level, then the cell is determined to exhibit a high likelihood of expressing an elevated level of PI3K, Akt, and/or MAPK activity upon being contacted with an mTORC1 inhibitor.
Other aspects of this invention provide methods for determining mTOR kinase activity in a cell by determining the level of phosphorylation of a phosphorylation site disclosed in Table 1, 2, 3, 7, 8, or 11 in the cell, and comparing the level of phosphorylation to a reference level, wherein if the level of phosphorylation is higher than the reference level, then the cell is determined to exhibit an elevated level of mTOR kinase activity, or if the level of phosphorylation is equal or lower than the reference level, then the cell is determined to not exhibit an elevated level of mTOR kinase activity. In certain embodiments, if a phosphorylation site determined to exhibit a higher level of phosphorylation in the cell as compared to the reference level is a class I phosphorylation site, then the cell is determined to exhibit an elevated level of rapamycin-sensitive mTORC1 activity. In certain embodiments, if a phosphorylation site determined to exhibit a higher level of phosphorylation in the cell as compared to the reference level is a class II phosphorylation site, then the cell is determined to exhibit an elevated level of rapamycin-insensitive mTORC1 and/or mTORC2 activity. In some embodiments, if a phosphorylation site determined to exhibit a higher level of phosphorylation in the cell as compared to the reference level is a class III phosphorylation site, then the cell is determined to exhibit an elevated level of rapamycin-sensitive mTOR activity, or an elevated level of rapamycin-insensitive mTOR activity, or both.
In some embodiments, the methods provided herein further comprise selecting a method of treatment of the subject based on the level of phosphorylation of the phosphorylation site. In some embodiments, if the cell is determined to exhibit an elevated level of mTOR kinase activity, then a method of treatment is selected that includes administering an effective amount of an mTOR kinase inhibitor to the subject. In some embodiments, if the cell is determined to not exhibit an elevated level of mTOR kinase activity, then a method of treatment is selected that does not include administering an mTOR kinase inhibitor. In some embodiments, if the cell is determined to exhibit an elevated level of rapamycin-sensitive mTORC1 activity, then a method of treatment is selected that comprises administering an effective amount of an inhibitor of rapamycin-sensitive mTORC1 kinase activity (e.g., rapamycin or a rapamycin analog) to the subject. In some embodiments, if the cell is determined to exhibit an elevated level of rapamycin-insensitive mTORC1 and/or mTORC2 activity, then a method of treatment is selected that comprises administering an effective amount of an inhibitor of rapamycin-insensitive mTORC1 kinase activity to the subject. In some embodiments, if the cell is determined to exhibit an elevated level of rapamycin-sensitive mTOR kinase activity, or an elevated level of rapamycin-insensitive mTOR kinase activity, or both, then a method of treatment is selected that comprises administering an effective amount of an inhibitor of rapamycin-sensitive and rapamycin-insensitive mTOR kinase activity or a combination of an inhibitor of rapamycin-sensitive mTOR kinase activity and an inhibitor of rapamycin-insensitive mTOR kinase activity to the subject.
In some embodiments, the method of treatment further comprises administering to the subject an effective amount of a compound that stabilizes Grb10 or that inhibits the degradation of Grb10. In some embodiments, the method of treatment further comprises administering an effective amount of an inhibitor of PI3K, Akt, or MAPK to the subject.
Some aspects of this invention provide a phosphoproteomics array that includes a plurality of phosphosensitive antibodies or antibody fragments each of which specifically binds to phosphorylation site disclosed in Table 1, 2, 3, 7, 8, or 11. In some aspects, the invention also provides a method of using a phosphoproteomics array to determine mTOR activity in a cell by contacting a proteinaceous sample derived from a cell, for example, from a cancer cell, with a phosphoproteomics array as described herein under conditions suitable for a protein expressed in the cell to bind to an antibody or antibody fragment of the array, determining the level of phosphorylated protein bound to an antibody or antibody fragment of the array, and comparing the level of phosphorylated protein bound to an antibody or antibody fragment of the array to a reference level. If the level phosphorylated protein in the sample derived from the cell is higher than the reference level, then the cell is determined to exhibit an elevated level of mTOR kinase activity.
In other aspects, the invention provides methods of identifying an mTOR kinase inhibitor by contacting an mTOR kinase with a polypeptide with a phosphorylation site disclosed in Table 1, 2, 3, 7, 8, or 11 under conditions suitable for the mTOR kinase to phosphorylate the phosphorylation site in the presence of a candidate agent, determining the level of phosphorylation at the phosphorylation site, and comparing the level of phosphorylation to a reference level. If the level of phosphorylation is lower than the reference level, then the candidate agent is identified as an mTOR kinase inhibitor.
In some embodiments involving a phosphorylation site disclosed in Table 1, 2, 3, 7, 8, or 11, the phosphorylation site is not a phosphorylation site previously known to be an mTOR target.
The subject matter of this application may involve, in some cases, interrelated products, alternative solutions to a particular problem, and/or a plurality of different uses of a single method or agent. Other advantages, features, and uses of the invention will become apparent from the following detailed description of non-limiting embodiments of the invention when considered in conjunction with the accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Sample preparation and data analysis for quantitative phosphorylation profiling of mTOR signaling. (A) Schematics of the two quantitative mass spectrometry experiments are shown with a plot highlighting the distribution of phosphopeptides identified in each screen. See data summary in Table 5. Note that most of the phosphopeptides have a ratio of 1:1 between the light and heavy populations and hence have a value close to 0 on a Log₂axis. Proteins with downregulated phosphorylation in each screen are highlighted in the red box. (B) Typical quantitative MS and MS/MS spectra in which LS*SLRAS*TSKSESSQK (SEQ ID NO: 1) from ribosomal protein S6 (S235 and S240) are identified as a rapamycin-sensitive phosphopeptide. Note the light and heavy peptides differ by 26 Da, corresponding to 2 labeled Lys and 1 labeled Arg in this particular peptide. Sequence in lower panel: SEQ ID NO: 1722. (C) Quantitative differences between the rapamycin-sensitive and -insensitive mTOR downstream phosphorylation events. Phosphopeptides identified in both screens were extracted and their corresponding Log₂ratios (fold-changes) were plotted. (D) The top ten pathways enriched in the down-regulated phospho-proteins identified in the rapamycin (Rapa) screen.

FIG. 2. Phosphorylation of Grb10 at S501/S503 is sensitive to rapamycin inhibition. (A) Identification of a doubly-phosphorylated rapamycin-sensitive Grb10 peptide (MNILSS*QS*PLHPSTLNAVIHR, SEQ ID NO: 2, * indicates phosphorylation sites) corresponding to S501/S503. (B) Evolutionary conservation of Grb10 S501/S503 among vertebrate species. (C) Phosphorylation of Grb10 at S501/S503 shows rapamycin sensitivity in vivo. TSC2−/− cells were starved for serum and treated with 20 nM rapamycin for the indicated times. Sequences, from top to bottom: SEQ ID NO: 1723 to SEQ ID NO: 1727, respectively. (D) Phosphorylation of Grb10 at S501/501 is sensitive to amino acids withdrawal. TSC2−/− cells were starved in DMEM overnight and then transferred to D-PBS for the indicated times. (E) Phosphorylation of Grb10 at S501/S503 is not affected by the pan-kinase inhibitor, staurosporine. TSC2−/− cells were starved for serum and treated with either 100 nM staurosporine or Ku-0063794 at the indicated concentrations for two hours. (F) Grb10 phosphorylation is increased upon growth factor stimulation. Wild type (WT) mouse embryonic fibroblasts (MEFs) were starved for serum overnight and then were stimulated with insulin (100 nM) or serum (10%) for 15 min. The cells were preincubated with the indicated compounds for two hours. AktVIII (1 μM) is a specific inhibitor of Akt whereas AZD (AZD6244, 5 μM) specifically inhibits MEK. Rapamycin (rapa) was used at 20 nM. (G) Grb10 phosphorylation at S501/S503 is sensitive to various mTOR kinase inhibitors. TSC2−/− cells were serum-starved and treated with the indicated compounds for two hours. The concentrations of the compounds were rapamycin 20 nM, LY (LY294002) 20 μM, BEZ235 (NVP-BEZ235) 500 nM, torin 100 nM, and pp 242 1 μM.

FIG. 3. mTOR-mediated Grb10 phosphorylation is important for Grb10 stability. (A) Grb10 interacts with raptor, but not rictor. HA-tagged Grb10 was co-transfected with Myc-raptor or Myc-rictor in HEK293T cells. Cells were lysed in lysis buffer A, and the lysates were subjected to immunoprecipitation using anti-HA antibody conjugated beads. Raptor and rictor were probed with an antibody against the Myc-tag. (B) Grb10 is phosphorylated by mTOR in vitro. Recombinant Grb10 was prepared from bacteria (the GST-fused Grb10 shows a molecular weight of 80 kDa) and was incubated with recombinant mTOR in vitro. Phosphorylation of Grb10 at S501/S503 was detected by using the phospho-specific antibody against these two sites. (C) Long-term rapamycin treatment leads to Grb10 degradation in TSC2−/− cells. Note that Grb10 protein expression levels inversely correlated with Akt activity. Grb10 is highly overexpressed in TSC2−/− cells. mRNA level was determined using quantitative RT-PCR based on three biological replicate experiments. (D) Knockdown of raptor in TSC2−/− cells decreased Grb10 protein level. Cells were starved overnight and the lysates were probed with the antibodies indicated. (E) Grb10 is highly overexpressed in TSC2−/− cells. (F) S501A/S503A mutant is unstable compared with the wild-type or the S501D/S503D mutant. The same amount of DNA was transfected into HEK293T cells. (G) Rapamycin failed to induce degradation of the S501D/S503D mutant. S501D/S503D mutant (DD) was stably expressed in TSC2−/− cells and cells were treated with 20 nM rapamycin for the indicated times. Endogenous Grb10 was detected using an antibody that preferentially recognizes mouse Grb10 whereas the Grb10 DD mutant (of human origin) was detected using an anti-HA antibody.

FIG. 4. Grb10 is involved in the feedback inhibition loop from mTORC1 to PI3K and MAPK and GRB10 mRNA expression is significantly down-regulated in many cancers and is negatively correlated with PTEN expression. (A) Knockdown of Grb10 in TSC2−/− cells resulted in PI3K and MAPK hyperactivation after insulin or IGF stimulation. (B) Knockdown of Grb10 in TSC2−/− cells protected cells against apoptosis. Grb10 knockdown and control cells were starved overnight and then treated with 100 nM staurosporine for 5 hrs to induce apoptosis. (C) Box plots indicating that GRB10 expression is significantly lower in many tumor types compared to their corresponding normal tissues. (Only the tumor types that showed significantly lower GRB10 expression in cancer vs. normal in at least three independent microarray datasets are included). (D) Heat maps indicating a strong negative correlation between GRB10 and PTEN expression in breast carcinomas and myelomas. Low levels of GRB10 expression rarely occurred in tumors that also showed low levels of PTEN expression. (E) Scatter plots comparing the expression levels of GRB10 and PTEN levels in the normal and tumor samples, collected overall from 6 different tissue types, where GRB10 was shown to be significantly down-regulated in cancer vs. normal. The negative correlation between GRB10 and PTEN expression is evident in the tumor (P<0.001) but not in the corresponding normal samples.

FIG. 5. Data analysis for the rapamycin screen and Ku-0063794 screen. (A) Phosphopeptides enrichment in the second biological replicate experiment of the rapamycin screen. Number of phospho- and nonphospho-peptides, in each SCX fraction was plotted. (B) Number of downregulated proteins in the two biological replicates of the rapamycin screen. (C) Ku-0063794 inhibits insulin-induced Akt phosphorylation. (D) Pathways enriched in the downregulated phospho-proteins identified in the Ku-0063794 screen. The top ten overrepresented pathways were shown. Analysis was performed using DAVID. See Huang D W, Sherman B T, Lempicki R A. Systematic and integrative analysis of large gene lists using DAVID Bioinformatics Resources. Nature Protoc. 2009; 4(1):44-57; and Dennis G Jr, Sherman B T, Hosack D A, Yang J, Gao W, Lane H C, Lempicki R A. DAVID: Database for Annotation, Visualization, and Integrated Discovery. Genome Biol. 2003; 4(5):P3; both incorporated herein by reference for disclosure of integrative analysis methods for large datasets.

FIG. 6. Examples of protein phosphorylation changes identified in the rapamycin and Ku-0063794 screens. Note that the light cells were treated with rapamycin and the heavy cells were treated with DMSO in the rapamycin screen (2^ndbiological replicate). For the Ku-0063794 screen, light cells were treated with rapamycin whereas the heavy cells were treated with a combination of rapamycin and Ku-0063794. (A) Phosphorylation changes of different sites on 4EBP in the rapamycin screen and Ku-0063794 screen. Phosphorylation of T36/T45 was partially responsive to rapamycin and was completely abolished as a result of Ku-0063794 treatment. In contrast, T70 phosphorylation is mTOR-independent (B) NRDG1 phosphorylation at S330/S333 was sensitive to Ku-0063794 but not rapamycin inhibition. (C) GSK3β phosphorylation at S9 decreased in both rapamycin and Ku-0063794 screen.

FIG. 7. Identification of the rapamycin-sensitive phosphorylation sites on mTOR (A) Intensities of an mTOR peptide ²⁴⁷¹AGTTVPES*HIS*FIGDGLVKPEALNKK²⁴⁹⁶(SEQ ID NO: 3, * indicates phosphorylation sites) from the rapamycin-treated (Light) and control (Heavy) TSC2−/− cells (B) Domain structure of mTOR and conservation of S2478 and S2481 across different species. Sequences, from top to bottom: SEQ ID NO: 1728 to SEQ ID NO: 1735, respectively. (C) Immunoblot experiments showing phospho-mTOR at S2481 is inhibited by acute rapamycin treatment. TSC2−/− cells were starved and then treated with 20 nM rapamycin for the indicated times. (D) mTOR kinase inhibitor pp 242 completely ablated mTOR phosphorylation at S2481. TSC2−/− cells were starved and were then treated with 1 pp 242 for two hours.

FIG. 8. Identification of rapamycin-sensitive phosphorylation sites on Grb10. (A) MS/MS experiments identified that phosphorylation of residues S501/S503 of Grb10 is strongly inhibited by rapamycin. (B) Phosphorylation at S455/S458 is rapamycin-insensitive. Sequences, from top to bottom: SEQ ID NO: 1736 and SEQ ID NO: 1737, respectively.

FIG. 9. Phosphorylation of Grb10 at S501/S503 regulates its stability. (A) An antibody showed specificity towards Grb10 phosphorylation at S501/S503. Preincubation of the antibody with the blocking antibody completely eliminated the immunoreactivity. (B) Further validation of the phospho-specific antibody raised against the Grb10 S501/S503 sites. Grb10 WT, S501A/S503A (AA) and S501D/S503D (DD) were transfected into HEK239T cells and were probed with the antibody. Note that this antibody detected wild type (WT) Grb10, but neither of the mutant proteins. (C) Treatment of TSC2−/− cells with various mTOR kinase inhibitors led to Grb10 degradation. (D) Both rapamycin and NVP-BEZ235 treatment led to a decreased level of Grb10 in TSC1−/− cells. (E) Grb10 is highly overexpressed in TSC1−/− cells compared with their wild-type counterparts. (F) rapamycin treatment of TSC2−/− cells stably expressing wild type (WT) Grb10 led to lower levels of both endogenous and exogenous Grb10.

FIG. 10. Grb10 is involved in the feedback loop from mTORC1 to PI3K. (A) Overexpression of Grb10 in HEK293 cells inhibited PI3K activation. W, wild type (WT) Grb10, A, AA mutant and D, DD mutant. Knockdown of Grb10 in TSC2−/− cells led to IRS hyperphosphorylation after insulin stimulation. (B) Overexpression of Grb10 in HEK239 cells suppressed IRS tyrosine phosphorylation and PI3K recruitment after insulin stimulation. HA-tagged Grb10 was transfected and IRS2 immunoprecipitates were analyzed. Phosphorylation at Y612 of IRS was detected. (C) HA-tagged Grb10 was transfected and p85 immunoprecipitates were analyzed. (D) Knockdown of Grb10 in TSC2−/− cells led to increased IRS Y612 phosphorylation upon insulin stimulation. (E) Knockdown of Grb10 in TSC2−/− cells protected these cells against etoposide-induced apoptosis. Cells were starved overnight and were treated with 100 μM etoposide for the indicated times.

DEFINITIONS

In the tables and the database, *, @, ̂ and # denote phosphorylation (Ser, Thr and Tyr), heavy Lys, heavy Arg and oxidation (Met), respectively. International Protein Index (IPI) reference numbers are given in some of the tables and the database provided herein. The IPI database can be accessed at the European Bioinformatics Institute homepage (www.ebi.ac.uk/), for example, at (www.ebi.ac.uk/IPI/IPIhelp.html). Each IPI database entry provided by accession number is incorporated herein by reference for disclosure of the respective proteins amino acid sequence and accompanying protein information.
The term “antibody,” as used herein, refers to an immunoglobulin, whether natural or wholly or partially synthetically produced. All derivatives thereof which maintain specific binding ability are also included in the term. The term also covers any protein having a binding domain which is homologous or largely homologous to an immunoglobulin binding domain. These proteins may be derived from natural sources, or partly or wholly synthetically produced. An antibody may be monoclonal or polyclonal. The antibody may be a member of any immunoglobulin class, including any of the human classes:IgG, IgM, IgA, IgD, and IgE. Derivatives of the IgG class, however, are preferred in the present invention.
The term “antibody fragment,” as used herein, refers to any derivative of an antibody which is less than full-length. Preferably, the antibody fragment retains at least a significant portion of the full-length antibody's specific binding ability. Examples of antibody fragments include, but are not limited to, Fab, Fab′, F(ab′)₂, scFv, Fv, dsFv, diabody, and Fd fragments. The antibody fragment may be produced by any means. For instance, the antibody fragment may be enzymatically or chemically produced by fragmentation of an intact antibody or it may be recombinantly produced from a gene encoding the partial antibody sequence. Alternatively, the antibody fragment may be wholly or partially synthetically produced. The antibody fragment may optionally be a single chain antibody fragment. Alternatively, the fragment may comprise multiple chains which are linked together, for instance, by disulfide linkages. The fragment may also optionally be a multimolecular complex. A functional antibody fragment will typically comprise at least about 50 amino acids and more typically will comprise at least about 200 amino acids.
Single-chain Fvs (scFvs) are recombinant antibody fragments consisting of only the variable light chain (V_L) and variable heavy chain (V_H) covalently connected to one another by a polypeptide linker. Either V_Lor V_Hmay be the NH₂-terminal domain. The polypeptide linker may be of variable length and composition so long as the two variable domains are bridged without serious steric interference. Typically, the linkers are comprised primarily of stretches of glycine and serine residues with some glutamic acid or lysine residues interspersed for solubility.
Diabodies are dimeric scFvs. The components of diabodies typically have shorter peptide linkers than most scFvs, and they show a preference for associating as dimers.
An Fv fragment is an antibody fragment which consists of one V_Hand one V_Ldomain held together by noncovalent interactions. The term dsFv is used herein to refer to an Fv with an engineered intermolecular disulfide bond to stabilize the V_H-V_Lpair.
A F(ab′)₂fragment is an antibody fragment essentially equivalent to that obtained from immunoglobulins (typically IgG) by digestion with an enzyme pepsin at pH 4.0-4.5. The fragment may be recombinantly produced.
A Fab fragment is an antibody fragment essentially equivalent to that obtained by reduction of the disulfide bridge or bridges joining the two heavy chain pieces in the F(ab′)₂fragment. The Fab′ fragment may be recombinantly produced.
A Fab fragment is an antibody fragment essentially equivalent to that obtained by digestion of immunoglobulins (typically IgG) with the enzyme papain. The Fab fragment may be recombinantly produced. The heavy chain segment of the Fab fragment is the Fd piece.
The term “binding agent”, as used herein, refers to an agent binding a target molecule, for example, a polypeptide comprising a phosphorylation site provided herein, with high specificity. Examples of binding agents are antibodies, antibody fragments, aptamers, and adnectins.
The term “phosphosensitive”, as used herein in the context of a binding agent, refers to a binding agent that specifically binds to a phosphorylation site, for example, a phosphorylation site provided herein, in either the phosphorylated or non-phosphorylated state. In some embodiments, a phosphosensitive binding agent provided herein binds to the phosphorylation site in its phosphorylated state, but does not significantly bind the phosphorylation site in a non-phosphorylated state. Phosphosensitive binding agents, for example, phosphosensitive antibodies or antibody fragments, accordingly, allow for the detection of phosphorylation at a specific phosphorylation site.
The term “cancer”, as used herein, refers to a malignant neoplastic disease. Most cancers are characterized by hyperproliferation of a cell population. In some embodiments, a cancer manifests as a solid tumor. In some embodiments, a cancer manifests as a liquid tumor. Non-limiting examples of cancers include carcinomas (derived from epithelial cells, e.g., some forms of breast, prostate, lung and colon cancer), sarcomas (derived from connective tissue or mesenchymal cells), lymphoma and leukemia (derived from hematopoietic cells) and seminomas (derived from germ cells).
The term “cancer cell”, as used herein, refers to a malignant neoplastic cell. In some embodiments, a cancer cell is part of a neoplastic cell population. In some embodiments, a cancer cell is a cell of a solid tumor. In some embodiments, a cancer cell is a cell of a liquid tumor. In some embodiments, a cancer cell carries a mutation that affects regulation of cell cycle control. In some embodiments, a cancer cell is a cell obtained from a tumor in a subject.
The term “candidate agent”, as used herein refers to a molecule to be tested for a specific property, for example, for its ability to inhibit mTOR kinase activity. In some embodiments, a candidate agent is a small molecule. In some embodiments, a candidate agent is a polypeptide or protein. In some embodiments, a candidate agent is a binding agent. In some embodiments, a candidate agent is a nucleic acid.
The term “determining a level of expression”, as used herein, refers to performing an assay to determine the level of a gene product expressed in a cell or tissue, for example, in a cancer cell or tumor tissue. In some embodiments, the assay includes obtaining a cell from a subject, for example, by biopsy. In some embodiments, the gene product is a transcript, for example, an mRNA. In other embodiments, the gene product is a protein, for example, an mTOR substrate disclosed herein, or a protein comprising a phosphorylation site disclosed herein. In some embodiments, the gene product is a protein that is phosphorylated at a specific phosphorylation site. In some embodiments, the gene product is a protein that is not phosphorylated at a specific phosphorylation site. Methods, assays, and reagents to determine the level of a gene product in a cell or tissue are described herein and are well known to those of skill in the art. See, for example, Sambrook et al., Molecular Cloning, second edition, Cold Spring Harbor Laboratory, Plainview, N.Y.; (1989), or Ausubel et al., Current Protocols in Molecular Biology, Current Protocols (1989), and DNA Cloning: A Practical Approach, Volumes I and II (ed. D. N. Glover) IREL Press, Oxford, (1985); each of which is incorporated herein by reference.
Methods to determine transcript levels include, for example, RT-PCR, northern blot, in situ hybridization, microarray assays, and massive parallel sequencing assays. Methods to determine protein levels include, for example, western blot, immunohistochemistry, ELISA, protein array assays, and mass spectrometry.
The term “high risk of expressing an elevated level of PI3K, Akt, and/or MAPK activity”, as used herein in the context of cells, refers to a condition in a cell that is likely to result in upregulation of PI3K, Akt, and/or MAPK kinase activity, when the cell is contacted with a therapeutic agent, for example, with an mTOR inhibitor. The term “risk” is used interchangeably with the term “likelihood” in this context. Such risk can be conferred through feedback-relief triggered by the therapeutic target. For example, in some embodiments, a cell, for example, a cell derived from a tumor in a subject, is determined to have a high likelihood of expressing an elevated level of PI3K, Akt, and/or MAPK, if it is established that the cell highly expresses a feedback-mediator, for example, phosphorylated Grb10, that limits PI3K, Akt, and/or MAPK expression. If the feedback-mediator is a known substrate of an oncogenic kinase, for example, if the feedback-mediator is the mTOR kinase substrate Grb10, then the cell is likely to exhibit feedback relief upon administration of an inhibitor of the kinase, for example, an mTOR kinase inhibitor, which, in turn, may increase the cell's proliferation and/or survival capacity.
The term “inhibitor” as used herein in the context of kinases, refers to a molecule that inhibits the activity of a kinase. In some embodiments, an inhibitor diminishes the catalytic activity of a kinase. In some embodiments, an inhibitor abolishes the catalytic activity of a kinase. In some embodiments, the inhibitor is a small molecule. In some embodiments, the inhibitor is a nucleic acid or a polypeptide. In some embodiments, the inhibitor is a binding agent. In some embodiments, a kinase inhibitor effects inhibition by down-regulating expression of the kinase. In some embodiments, a kinase inhibitor effects inhibition by binding the kinase and interfering with the kinase reaction. In some embodiments, a kinase inhibitor is an allosteric kinase inhibitor. The term “allosteric kinase inhibitor” refers to a kinase inhibitor, for example, a small molecule, that binds its target kinase, wherein the binding of the inhibitor results in an allosteric change in the kinase molecule, leading to diminished kinase activity. Allosteric changes leading to diminished kinase activity can be changes resulting in a reduction of the kinase's ability to bind a substrate molecule, or changes resulting in a reduction of the kinases ability to transfer a phosphate group to a substrate molecule. In some embodiments, the kinase inhibitor is a catalytic kinase inhibitor. In some embodiments, a catalytic inhibitor, which itself cannot partake in a kinase reaction, competes with a kinase substrate for binding. In some embodiments, a catalytic kinase inhibitor is an ATP-competitive inhibitor.
The term “effective amount”, as used herein in the context of kinase inhibitors, refers to an amount of kinase inhibitor sufficient to achieve a measurable inhibition of kinase activity, or an amount of kinase inhibitor sufficient to achieve a clinically desirable outcome. For example, an effective amount of the mTOR inhibitor rapamycin is, in some embodiments, an amount of rapamycin that, when administered to a cell or tissue, results in a measurable decrease in mTOR kinase activity in the cell or tissue. In some embodiments, an effective amount of a kinase inhibitor is an amount that is effective to reduce the activity of the target kinase in a cell or tissue to less than 75%, less than 50%, less than 40%, less than 30%, less than 25%, less than 20%, less than 10%, less than 5%, less than 2.5%, less than 2%, or less than 1% of the activity in the cell or tissue not treated with the inhibitor. Assays for measuring kinase activity are well known in the art and described in more detail elsewhere herein. In some embodiments, an effective amount of a kinase inhibitor, for example, of rapamycin, is an amount that, when administered to a subject, for example, a subject having a cancer with elevated mTOR activity, results in a clinically desirable outcome. In some embodiments, a clinically desirable outcome is reversal of a disease, for example, a decrease in neoplastic or malignant cell number, tumor size, or cell proliferation rate, for example, in a subject having a proliferative disease, e.g., a neoplastic disease or a cancer, or a delay in the progression of a disease, for example a progression from one tumor stage to the next, or from a benign to a malignant neoplastic disease.
The term “mTOR kinase inhibitor” as used herein, refers to a molecule inhibiting a kinase activity of the mTOR kinase. Examples of mTOR kinase inhibitors are mTORC1 kinase inhibitors and mTORC1/2 inhibitors. Examples of mTORC1 kinase inhibitors are rapamycin and rapamycin analogs (e.g., Ridaforolimus, Sirolimus or Everolimus). Examples of mTORC1/2 inhibitors are PP242, PP30, AZD8055, OSI-027, WYE354, INK-128, XL388, and torin1. Further examples of mTOR inhibitors are inhibitors that target mTOR and an additional kinase or additional kinases, for example, dual PI3K/mTOR kinase inhibitors (e.g., NVP-BEZ235, BGT-226, XL-765, GSK2126458, or SF1126).
The term “PI3K kinase inhibitor”, as used herein, refers to a molecule that inhibits PI3K kinase activity. Examples of PI3K inhibitors are ZSTK474, TGX221, GDC0941, LY294002, XL147, PX147, BKM120, GSK 615, CAL101, and PX-866.
The term “mutation”, as used herein, refers to a change in a gene sequence, for example, a deletion, insertion, inversion, transposition, or substitution. In some embodiments, the mutation results in a change of the expression level of the gene product encoded by the respective gene. In some embodiments, a mutation is a mutation in a gene involved in an mTOR signaling pathway, for example, a gene encoding a protein that regulates, directly or indirectly, mTOR kinase activity. Examples of genes involved in an mTOR signaling pathway are Ras, Raf, MAPK, RSK, receptor tyrosine kinases, PI3K (Phosphoinositide 3-kinase), PTEN (phosphatase and tensin homolog), Akt (Protein Kinase B), TSC1/2 (Tuberous sclerosis protein 1, Tuberous sclerosis protein 2, respectively), MEK (Dual specificity mitogen-activated protein kinase kinase 1, MAPK21), LKB (, and NF2 (Neurofibromatosis 2). Methods to determine whether a cell carries a mutation in a gene are well known to those of skill in the related arts.
The term “phosphoproteomic profile”, as used herein, refers to a dataset comprising information regarding the level of phosphorylation of a plurality of phosphorylation sites in a biological sample, for example, a proteinaceous sample derived from a cell or tissue sample. Phosphoproteomic profiles of multiple samples can be compared and similarities and dissimilarities in such profiles can be detected and quantified by methods well known to those of skill in the art, including, but not limited to, supervised and non-supervised learning, hierarchical clustering, nearest neighbor analysis. In some embodiments, a phosphoproteomics profile of a clinical sample, for example, of a sample derived from a malignant cell or tissue sample of a subject is compared to a reference sample from healthy cells or tissue, for example, to determine aberrations in protein phosphorylation in the malignant cell or tissue sample. In some embodiments, a phosphoproteomics profile of a clinical sample at issue is compared to phosphoproteomics profiles of clinical samples of known character, for example, to classify the clinical sample at issue. Phosphoproteomic profiles can be classified by methods well known in the art, including the building and application of predictors for the classification.
The term “phosphorylation level”, as used herein, refers to the proportion of phosphorylated polypeptides carrying a certain phosphorylation site in a sample relation to all polypeptides carrying the phosphorylation site in the sample. For example, if the total number of polypeptides carrying a specific phosphorylation site in a sample is 10, and 3 of these proteins are phosphorylated at that site, while 7 are not, then the phosphorylation level of this phosphorylation site in the sample would be 30%. Phosphorylation levels can be detected and quantified by methods known to those in the art, for example, by protein detection using phosphosensitive binding agents, e.g., phosphosensitive antibodies or antibody fragments. Most protein quantitation methods known to those of skill in the art do not allow for precise molecule counting, so phosphorylation levels are often approximated based on estimations of the number or fractions of phosphorylated proteins in a sample, for example, after western blot or protein microarray analysis using phosphosensitive antibodies or antibody fragments.
The term “phosphorylation site”, as used herein, refers to an amino acid residue within an amino acid sequence, or motif, that can be phosphorylated, for example, by a kinase targeting the respective site. A phosphorylation site is a substrate of a kinase if it can be phosphorylated by that kinase. For example, 5421 and S432 are phosphorylation sites in Grb10 isoform 3 within the motif MSNILSS*QS*PLHPSTLNAVIHR (SEQ ID NO: 4), and are both mTOR substrates, as described elsewhere herein. mTOR phosphorylation sites can further be classified into three classes, as used herein. The term “class I phosphorylation site”, as used herein, refers to a phosphorylation site that is targeted by rapamycin-sensitive mTORC1 activity. The term “class II phosphorylation site,” as used herein, refers to a phosphorylation site that is targeted by rapamycin-insensitive mTORC1 and/or mTORC2 activity. The term “class III phosphorylation site,” as used herein, refers to a phosphorylation site that is targeted by rapamycin-sensitive mTOR activity, rapamycin-insensitive mTOR activity, or both.
The term “proliferative disease,” as used herein, refers to any disease in which cell or tissue homeostasis is disturbed in that a cell or cell population exhibits an abnormally elevated proliferation rate. Proliferative diseases include hyperproliferative diseases, such as pre-neoplastic hyperplastic conditions and neoplastic diseases. Neoplastic diseases are characterized by an abnormal proliferation of cells and include both benign and malignant neoplasias. Malignant neoplasia is also referred to a s cancer.
The term “reference”, interchangeably used with the term “control” herein, refers to a value, sample, or parameter that serves as a baseline for comparing a value, sample, or parameter of interest to. The use of a reference can be of value in many methods that allow for the detection of absolute levels, for example, of expression, phosphorylation, or protein binding, and is essential in methods that yield semi-quantitative or relative results. Comparing an assay result obtained for a cell or tissue in question, for example, a cell or tissue obtained from a tumor in a subject, to a reference result allows a determination whether the result is abnormal. Depending on their nature, abnormal results can support the inference of specific molecular or cellular aberrations and, in some embodiments, a selection of a course of treatment over another. In some embodiments, a reference value is obtained from cells of the same cell type or the same tissue of origin as the cell in question. For example, in some embodiments, a diseased and a healthy cell is obtained from a subject, and the cells are assayed by methods provided herein in parallel. The value observed in the healthy cell, for example, a level of phosphorylation of a protein then typically serves as the reference level to which the level observed in the cell in question is compared. In other embodiments, the reference level is an average level observed or expected in normal cells. In some embodiments, the reference level is a range typically observed in healthy cells. A suitable reference depends, of course, on the type of assay and sample involved. A suitable reference for a given assay or sample will readily be apparent to those of skill in the art. The following list of exemplary references is for illustration only, since the invention is not limited in this respect.
In some embodiments, a suitable reference level, for example, a suitable reference phosphorylation or expression level, is a level observed or expected in a healthy cell or tissue of the same type as the cell or tissue in question. In some embodiments, for example, in some embodiments where a tumor biopsy is assayed, a suitable reference level is a level observed or expected in healthy cells or tissue of the same tissue the tumor originated from, or in cells or tissue adjacent to the tumor. In some embodiments, for example, in some embodiments where a cell is assayed, a reference cell is a healthy cell that is of the same cell type or tissue of origin as the cell in question. In some embodiments, a reference cell is a cell exhibiting normal mTOR kinase activity. In some embodiments, a reference cell is a malignant cell of known phenotype, for example, a malignant cell known to exhibit elevated mTOR kinase activity, a cell known to exhibit elevated mTORC1 kinase activity, or a cell known to exhibit elevated mTORC1/2 kinase activity. In some embodiments, a reference cell is a malignant cell of known phenotype, for example, a rapamycin-sensitive cancer cell or a rapamycin-sensitive cancer cell.
In some embodiments, a suitable reference level is an average level calculated or approximated from historic data. In some embodiments, a reference level is a level obtained from a reference cell or tissue assayed in parallel to the cell or tissue in question. In some embodiments, a level obtained from a cell or tissue sample is determined to be different (higher or lower) than the reference level, if it is statistically significantly different (higher or lower) than the reference level. In some embodiments, a level obtained from a cell or tissue sample is determined to be higher than the reference level, if the level is at least about 1.25-fold, at least about 1.5-fold, at least about 2-fold, at least about 3-fold, at least about 5-fold, or at least about 10-fold the reference level. In some embodiments, a level obtained from a cell or tissue sample is determined to be lower than the reference level, if it is less than about 0.75-fold, less than about 0.70-fold, less than about 0.60-fold, less than about 0.50-fold, less than about 0.40-fold, less than about 0.30-fold, less than about 0.25-fold, less than about 0.10-fold, less than about 0.05-fold, or less than about 0.01-fold the reference level. In some embodiments, a level obtained from a cell or tissue is determined to be substantially similar to the reference level, if it is not statistically significantly different from the reference level. In some embodiments, a level obtained from a cell or tissue is determined to be substantially similar to a reference level, if it is within the range of 0.75-fold to 1.25-fold of the reference level. In some embodiments, a profile comprising a plurality of levels obtained from a cell or tissue sample is determined to be substantially similar to a reference profile, if the profiles cluster together in a clustering analysis. In some embodiments, a multi-value profile obtained from a cell or tissue is determined to be substantially similar to a reference profile, if the correlation coefficient between the profiles is at least 0.75, at least 0.8, at least 0.85, at least 0.9, at least 0.91, at least 0.92, at least 0.93, at least 0.94, at least 0.95, at least 0.96, at least 0.97, at least 0.98, or at least 0.99.
The term “sample”, as used herein, refers to a biological sample. A biological sample typically comprises a cell or tissue, or biological material derived from a cell or tissue. Examples of biological samples are cell samples and tissue samples obtained from a subject, for example, from a subject having a cancer, cell or tissue cultures, and extracts or preparations obtained from any such samples, for example, protein extracts, isolated proteins or isolated nucleic acids obtained from such samples.
A “proteinaceous” sample is a sample comprising proteins or fragments of proteins. In some embodiments, a proteinaceous sample is a sample derived from a cell that includes proteins or fragments of proteins expressed in the cell. In some embodiments, the sample includes a lysed cell. In some embodiments, the sample includes proteins isolated from a cell. In some embodiments, the sample includes an isolated, enriched, or purified protein fraction from a cell, for example, a protein fraction obtained by methods described herein or known to those of skill in the art to separate nucleic acids, carbohydrates, and/or lipids from proteins or fractions of proteins. In some embodiments, the sample is prepared under conditions suitable for obtaining native proteins from a cell. In some embodiments, the sample is prepared under conditions that do not significantly affect native protein phosphorylation patterns. In some embodiments, the sample is prepared under conditions that do not significantly affect the antigenic structure of a phosphorylation site identified herein, for example, a phosphorylation site identified herein to be a target of mTOR kinase activity. In some embodiments, the sample is prepared under conditions suitable for obtaining denatured or fragmented proteins.
The term “subject,” as used herein, refers to an individual mammal. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human mammal. In some embodiments, the subject is a non-human primate. In some embodiments, the subject is a rodent. In some embodiments, the subject is a sheep, a goat, a cattle, a cat, or a dog. In some embodiments, the subject is a subject diagnosed to have a cancer. In some embodiments, the subject is a subject diagnosed to have a cancer that exhibits an elevated level of mTOR kinase activity. In some embodiments, the subject is a subject not diagnosed with a cancer.
The term “target,” as used herein in the context of kinases, refers to a substrate of a kinase. The term can, accordingly, refer to a protein (e.g., Grb10 is a target of mTOR kinase), to an amino acid sequence comprising a phosphorylation site (e.g., MNILSS*QS*PLHPSTLNAVIHR (SEQ ID NO: 5) or MNILGS*QS*PLHPSTLSTVIHR (SEQ ID NO: 6) are target sequences of mTOR), or to an amino acid residue (e.g. S421 and S423 of mouse Grb10, isoform 3, are targets of mTOR).
The term “tumor,” as used herein, refers to a neoplasm or a solid lesion formed by neoplastic cells. A tumor can be benign, pre-malignant, or malignant. In some embodiments, the tumor is a malignant tumor.
The term “treatment”, interchangeably used herein with the term “therapy,” as used herein, refers to a clinical intervention aimed to prevent or to ameliorate a disease or condition in a subject. In some embodiments, a treatment is aimed to ameliorate an existing condition, for example, a cancer in a subject. In some embodiments, a treatment is aimed to prevent a condition from occurring or from recurring. For example, in some embodiments, an mTOR kinase inhibitor is administered to a subject having a cancer exhibiting an elevated level of mTOR activity in order to inhibit cell proliferation in the malignant cells. For another example, in some embodiments, an mTOR kinase inhibitor is administered to a subject having a cancer exhibiting an elevated level of mTOR activity after a malignant tumor has been removed from the subject, in order to prevent or delay recurrence of the disease. The foregoing examples are non-limiting, and those of skill in the art will readily envision further scenarios of treatment as the invention is not limited in this respect. Some embodiments include a choice of treatment, referring to a selection of a clinical intervention from a number of alternatives, or to a design of a clinical intervention to meet the specific needs of an individual subject. In some embodiments, a choice of treatment involves the design of a personalized therapeutic approach for a subject having a cancer exhibiting elevated mTOR activity based on the results from diagnostic methods provided herein. For example, in some embodiments, a choice of treatment includes administering to a subject having cancer a specific mTOR inhibitor in combination with an additional kinase inhibitor targeting an mTOR feedback-inhibited pathway based on a determination that cancer cells in the subject exhibit high mTOR activity and elevated levels of a feedback-inhibitor targeted by mTOR in order to avoid feedback relief. In some embodiments, a choice of treatment includes the determination of an appropriate treatment. Some embodiments further include carrying out the selected treatment.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

This invention relates to the identification of proteins and sites on those proteins that are phosphorylated by mTOR kinase. Some aspects of this invention relate to the surprising discovery that some proteins that are substrates of mTOR kinase regulate mTOR upstream signaling pathways, thus providing feedback regulatory circuits. Some aspects of this invention relate to methods of classifying cells, for example, neoplastic cells or cancer cells, based on the phosphorylation status of mTOR targets. Some aspects of this invention relate to methods of personalized treatment of subjects having a proliferative disease, for example, a neoplastic disease or a cancer, exhibiting elevated mTOR signaling based on the phosphorylation status of mTOR targets in the diseased cells. Other aspects of this invention relate to methods and materials for the generation of phosphoproteomics profiles that include phosphorylation data for a plurality of mTOR targets. Some aspects of this invention relate to classification of cells or tissues, for example, tumor cells or tissues, based on phosphoproteomic data, for example, based on phosphoproteomic profiles of the cells or tissues. Some aspects of this invention relate to methods of identifying mTOR kinase inhibitors by identifying compounds able to interfere with mTOR kinase activity towards one or more of the mTOR targets provided herein. Some aspects of this invention relate to the identification of Grb10 as a target of mTOR kinase activity, and to methods for determining the level of mTOR activity in a cell based on Grb10 phosphorylation status. Some aspects of this invention relate to the surprising discovery that Grb10 is not only a target of mTOR kinase activity, but also an inhibitor of PI3K kinase activity. Since PI3K is a positive regulator of mTOR kinase activity, Grb10 has been identified as a mediator of mTOR feedback inhibition herein. Other aspects of this invention relate to the surprising discovery that Grb10 and PTEN mutations are mutually exclusive in certain types of tumor cells. Accordingly, some aspects of this invention relate to the surprising discovery that Grb10 is a tumor suppressor and that Grb10 phosphorylation is a useful biomarker for determining mTOR kinase activity in cells, and particularly in tumor cells. Other aspects of this invention relate to methods and reagents useful for determining the phosphorylation status of a protein or phosphorylation site identified to be an mTOR target herein. For example, some aspects of this invention provide phosphosensitive antibodies or antibody fragments specifically binding an mTOR kinase target site identified herein in either the phosphorylated or the non-phosphorylated state, thus allowing for a determination of the phosphorylation state of such a site in a cell.
mTOR Target Proteins and Sites
Identification of mTOR Target Sites
Some aspects of this invention relate to the identification of proteins and phosphorylation sites thereon as downstream targets of mTOR kinase activity. Targets of mTOR kinase activity are provided in Tables 1 and 2.
Classification of mTOR Targets
Some aspects of this invention relate to the classification of mTOR target phosphorylation sites provided herein as class I (targets of rapamycin-sensitive mTORC1 kinase signaling), class II (targets of rapamycin-insensitive mTORC1 or mTORC2 signaling), and class III (targets of both rapamycin-sensitive and rapamycin-insensitive mTOR signaling) phosphorylation sites.
This classification is useful to determine the type(s) of mTOR signaling exhibited by a cell or tissue and can support a classification of cells that are targeted for therapy, for example, of cancer cells. For example, in some embodiments, the phosphorylation status of an mTOR target is determined in a cancer cell obtained from a subject with a cancer exhibiting elevated mTOR kinase activity. Classification of the type of mTOR signaling can, in some embodiments, be a basis for the selection of an appropriate therapeutic approach. For example, if it is determined that a class I target of mTOR signaling is phosphorylated, then the elevated mTOR kinase activity is rapamycin-sensitive mTORC1 kinase activity. In some such embodiments, a method of treatment is then typically selected that includes administration of an inhibitor of rapamycin-sensitive mTORC1 signaling, for example, rapamycin or a rapamycin analog.
To give but another example: if it is determined that a class II target of mTOR signaling is phosphorylated, then the elevated mTOR kinase activity is rapamycin-insensitive mTORC1 or mTORC2 kinase activity. In some such embodiments, a method of treatment is then typically selected that includes administration of an mTORC1/2 inhibitor as provided herein. In some embodiments where class I targets of mTOR signaling are found to not be phosphorylated, a method of treatment is typically selected that does not include administration of an mTORC1 inhibitor.
To give a third example: if it is determined that a class III target of mTOR signaling is phosphorylated, then a method of treatment is selected that includes administration of an mTORC1 inhibitor and an mTORC1/2 inhibitor as provided herein. In some embodiments, a method of treatment that includes administering an mTORC1 inhibitor or an mTORC1/2 inhibitor alone may be inappropriate.
Further, some aspects of this invention provide methods to analyze gene ontology distributions in sets of mTOR targets, for example, in order to determine whether a specific signaling pathway is targeted by mTOR in a given cell, or a given therapeutic or experimental scenario.
Exemplary Proteins Identified in the Rapamycin Phosphoproteomic Screen as Targets of mTORC1 Signaling.
The phosphoproteomic screen described herein identified numerous proteins as mTOR targets, for example, as mTORC1 targets. As described in more detail elsewhere herein, the expression, expression level, phosphorylation, or phosphorylation level of one or more of such mTOR target proteins, for example, the proteins described in any of tables 1-3, or the level of phosphorylation of their respective phosphorylation sites, is employed, in some embodiments, as a biomarker for monitoring or diagnosing disease. In some embodiments, the mTOR target proteins disclosed herein, for example, the mTORC1 target proteins described in any of tables 1-3, are used in pharmaceutical screens as drug targets for the development of drugs modulating mTOR pathway downstream effects. Some exemplary mTORC1 target proteins identified herein are described in more detail below. These descriptions of exemplary mTOR target proteins are for illustration of some aspects of this disclosure and are not meant to limit the scope of the invention. Grb10, one of the mTORC1 protein targets identified in the phosphoproteomics screen, is described in more detail elsewhere herein. Briefly, Grb10, also known as growth factor receptor-bound protein 10, is stabilized by mTORC1-mediated phosphorylation, which, in turn, results in feedback inhibition of the PI3K and MAPK pathways. This is consistent with Grb10 expression being frequently downregulated in a variety of cancers. Grb10 is a tumor suppressor with relevance across a broad spectrum of cancer subtypes, and the identification of Grb10 as an mTORC1 substrate links mTORC1 activity to cancers with aberrantly low Grb10 expression or stability. Accordingly, some embodiments provide Grb10 phosphorylation as a biomarker for mTORC1 activity in a cell, tissue, or sample of interest. Methods of using Grb10 expression or phosphorylation assays to monitor mTORC1 activity in a cell, tissue, or sample of interest are also provided. For example, some embodiments provide a method of monitoring the efficacy of a drug on mTORC1 activity in a cell, tissue, or sample, by measuring Grb10 expression or phosphorylation levels. In some embodiments, such assessments are used to diagnose or monitor a disease associated with aberrant levels of mTORC1 activity, for example, cancer, or to adjust a drug dosage, for example, to the minimal dosage necessary to achieve a target mTORC1 activity level, or to choose a suitable drug modulating mTORC1 activity from a plurality of drugs, for example, the drug that shows the highest efficacy at a given dose.
Some of the mTORC1 protein targets identified herein are potential regulators of gene expression. For example, FOXK1 (Forkhead box protein K1), also known to those of skill in the art as MNF, or MNF (myocyte nuclear factor)-beta, is a winged-helix protein expressed selectively and transiently in myogenic precursor cells of the heart and skeletal muscles, and collaborates with proteins of the mammalian Sin3 (mSin3) family to repress transcription. Mutated forms of MNF-beta that fail to bind mSin3 are defective in transcriptional repression and in negative growth regulation, an overexpression phenotype revealed in oncogenic transformation assays. These phenotypic traits associated with MNF-beta dysfunction are consistent with the mTORC1 phenotype.
For another example, ZEB2 (Zinc finger E-box-binding homeobox 2) belongs to the ZEB family of zinc finger transcription factors, which are essential regulators of gene expression during normal embryonic development. ZEB proteins induce epithelial to mesenchymal transition (EMT), a process in which epithelial cells become migratory mesenchymal cells. E-cadherin is a major target gene of ZEB transcriptional repressors, and e-cadherin downregulation is considered a hallmark of EMT. In recent years, the involvement of the ZEB proteins in pathological contexts has been documented as well. For example, ZEB proteins play an important role in mediating Ras-induced EMT in breast epithelial cells. Mutations in ZEB encoding genes cause severe syndromic malformations, and are implicated in malignant tumor progression. Without wishing to be bound by theory, ZEB2 is believed to be a critical target in lymphangioleiomyomatosis (LAM) that represents the point of convergence of the mTORC1 and ERK-MAP kinase pathways that are critical to this disease.
For another example, NDRG3 is the downstream target of N-Myc. Phosphorylation of NDRG3 links mTOR signaling pathway to Myc activity as an oncogenic transcription factor.
Accordingly, some embodiments provide FOXK1, ZEB2, and/or NDRG3 phosphorylation as a biomarker for mTORC1 activity in a cell, tissue, or sample of interest. Methods of using FOXK1, ZEB2, and/or NDRG3 expression or phosphorylation assays and levels useful for monitoring mTORC1 activity in a cell, tissue, or sample of interest are also provided. For example, some embodiments provide a method of monitoring the efficacy of a drug on mTORC1 activity in a cell, tissue, or sample, by measuring FOXK1, ZEB2, and/or NDRG3 expression or phosphorylation levels. In some embodiments, such assessments are used to monitor or diagnose a disease associated with aberrant mTORC1 activity, as manifest by aberrant FOXK1, ZEB2, and/or NDRG3 phosphorylation level(s), and/or to adjust a drug dosage, for example, to the minimal dosage necessary to achieve a target mTORC I activity level, or to choose a suitable drug modulating mTORC1 activity from a plurality of drugs, for example, the drug that exhibits the highest efficacy at a given dose.
Some of the mTORC1 protein targets identified herein are regulators of mRNA synthesis, mRNA processing, and protein synthesis. This is consistent with the role mTORC1 activity plays in the control of cell growth, which, if improperly regulated, can contribute to tumor genesis and/or growth. For example, LARP1 binds mRNA in vitro via both the La motif and the LARP1 domain. LARP-1 also down-regulates the Ras-MAPK pathway. Accordingly, without wishing to be bound by any particular theory, LARP1 phosphorylation represents an mTORC1-dependent regulation of this feedback loop. The LARP-1 protein colocalizes with P bodies, which function in RNA degradation, and it is believed that the cluster of LARP-1 homologs functions to control the expression of key developmental regulators. Without wishing to be bound by any particular theory, some aspects of this invention provide that LARP-1 functions in P-bodies to attenuate the abundance of Ras-MAPK pathway-regulated mRNAs.
For another example, EDC3 is associated with an mRNA-decapping complex required for removal of the 5′-cap from mRNA prior to its degradation from the 5″-end.
For another example, SRPK2 is involved in SR protein phosphorylation, which influences other aspects of mRNA metabolism, such as splice site selection (alternative splicing), mRNA export, nonsense-mediated decay (NMD), and translation efficiency. Accordingly, some aspects of this disclosure provide that SRPK2 links mTOR/S6K signaling to SR protein activity.
Another example of an MTORC1 target identified herein that regulates mRNA synthesis and processing is CDK12-(CDC2-related kinase, arginine/serine-rich). CDK12 is involved in the regulation of alternative mRNA splicing. Without wishing to be bound by any particular theory, it is believed that, similar to SRPK2, CDK12 is also involved in regulating mRNA splicing by mTOR signaling. A CDK12-binding partner, cyclin-L1 (CCNL1), is also identified as an mTORC1 target protein herein. Without wishing to be bound by any particular theory, similar to SRPK and CDK12, CCNL1 may contribute to mRNA biogenesis and cell proliferation.
Accordingly, some embodiments provide LARP1, SRPK2, and/or CDK12 phosphorylation as a biomarker for mTORC1 activity in a cell, tissue, or sample of interest. Methods of using LARP1, SRPK2, and/or CDK12 expression or phosphorylation levels to monitor mTORC1 activity in a cell, tissue, or sample of interest are also provided. For example, some embodiments provide a method of monitoring the efficacy of a drug on mTORC1 activity in a cell, tissue, or sample, by measuring LARP1, SRPK2, and/or CDK12 expression or phosphorylation levels. In some embodiments, such assessments are used to monitor or diagnose a disease associated with aberrant mTORC1 activity, as manifest by aberrant LARP1, SRPK2, and/or CDK12 phosphorylation level(s), and/or to adjust a drug dosage, for example, to the minimal dosage necessary to achieve a target mTORC1 activity level, or to choose a suitable drug modulating mTORC1 activity from a plurality of drugs, for example, the drug that exhibits the highest efficacy at a given dose.
Another example of an mTORC1 target protein identified herein is MIB1, an E3 ubiquitin-protein ligase that has been reported to ubiquinate Notch, which leads to the degradation of Notch. Accordingly, some aspects of this disclosure provide that mTORC1 activity is linked to Notch signaling via phosphorylation of MIB1. Yet another example of an mTORC1 target protein identified herein is IBTK (Isoform 2 of Inhibitor of Bruton tyrosine kinase), which is a protein tyrosine kinase implicated in the primary immunodeficiency disease X-linked agammaglobulinemia (Bruton agammaglobulinemia). IBTK is activated upon binding to PIP3 generated as a result of PI3K activation. Without wishing to be bound by any particular theory, phosphorylation of IBTK affects IBTK's ability to inhibit PI3K kinase activity. Accordingly, some embodiments of this invention are based on the recognition that there is a link between the mTORC1 signaling system and the regulation of tyrosine phosphorylation.
Accordingly, some embodiments provide MIB1 and/or IBTK phosphorylation as a biomarker for mTORC1 activity in a cell, tissue, or sample of interest. Methods of using MIB1 and/or IBTK expression or phosphorylation assays to monitor mTORC1 activity in a cell, tissue, or sample of interest are also provided. For example, some embodiments provide a method of monitoring the efficacy of a drug on mTORC1 activity in a cell, tissue, or sample, by measuring MIB1 and/or IBTK expression or phosphorylation levels. In some embodiments, such assessments are used to monitor or diagnose a disease associated with aberrant mTORC1 activity, as manifested by aberrant MIB1 and/or IBTK phosphorylation level(s), and/or to adjust a drug dosage, for example, to the minimal dosage necessary to achieve a target mTORC1 activity level, or to choose a suitable drug modulating mTORC1 activity from a plurality of drugs, for example, the drug that exhibits the highest efficacy at a given dose.
Representative protein sequences of Grb10, FOXK1, ZEB2, NDRG3, LARP1, SRPK2, CDK12, MIB1, IBTK, and other proteins identified herein to be mTORC1 substrates, can be found under the respective database entries provided in the tables and the database provided herein, and, for example, in the NCBI database under accession numbers NP_—001171100.1 (growth factor receptor-bound protein 10 (Grb10) isoform 2, Mus musculus, SEQ ID NO: 1720), NP_—034475.2 (growth factor receptor-bound protein 10 (Grb10) isoform 1, Mus musculus, SEQ ID NO: 1721), NP_—001001550.1 (growth factor receptor-bound protein 10 (Grb10) isoform c, SEQ ID NO: 7), NP_—005302.3 (Grb10 isoform a, SEQ ID NO: 8) and NP_—001001549.1 (Grb10 isoform b, SEQ ID NO: 9); NP_—001032242.1 (FOXK1, SEQ ID NO: 10); NP_—001165124.1 and NP_—055610.1 (ZEB2, SEQ ID NO: 11 and SEQ ID NO: 12, respectively); NP_—071922.2 and NP_—114402.1 (NDRG3, SEQ ID NO: 13 and SEQ ID NO: 14, respectively); NP_—056130.2 (LARP1, SEQ ID NO: 15); NP_—872633.1 and NP_—872634.1 (SRPK2, SEQ ID NO: 16 and SEQ ID NO: 17, respectively); NP_—055898.1 and NP_—057591.2 (CDK12, SEQ ID NO: 18 and SEQ ID NO: 19, respectively); NP_—065825.1 (MIB1, SEQ ID NO: 20); and NP_—056340.2 (IBTK, SEQ ID NO: 21). These database entries are incorporated herein by reference for disclosure of representative sequences of the respective proteins.
Feedback Circuitry Involving mTOR Targets
Some aspects of this invention relate to the surprising discovery that some mTOR target proteins are involved in feedback loops of mTOR signaling, for example, by inhibiting upstream modulators of mTOR signaling, such as PI3K, Akt, and MAPK, after being phosphorylated as a result of mTOR kinase activity. Some aspects of this invention relate to the surprising discovery that relief of feedback inhibition of cellular pro-survival, pro-proliferation signaling by mTOR downstream targets, for example, by Grb10, can contribute to rapamycin resistance or even clinically detrimental outcome in the treatment of neoplastic disease (e.g., cancer) with mTOR inhibitors. Some aspects of this invention relate to the identification, for the first time, of Grb10 as a target of mTOR kinase activity. Grb10, growth factor receptor-bound protein 10, is a protein well known to those of skill in the art. Representative protein sequences of Grb10 can be found under the respective database entries provided in the tables and the database provided herein, and, for example, in the NCBI database under accession numbers NP_—001001550.1 (growth factor receptor-bound protein 10 isoform c), NP_—005302.3 (isoform a) and NP_—001001549.1 (isoform b). These database entries are incorporated by reference for disclosure of representative Grb10 sequences.

Feedback Inhibition of PI3K by Grb10

Some aspects of this invention relate to the surprising discovery that Grb10 is not only a target of mTOR kinase activity, but also a mediator of mTOR feedback inhibition and, thus, a tumor suppressor gene. As described in more detail elsewhere herein, mTOR-mediated phosphorylation of Grb10 results in stabilization and/or prevention of degradation of Grb10. Grb10, in turn, inhibits PI3K kinase activity. PI3K kinase activity targets mTOR and activates mTOR signaling. Accordingly, as provided by some aspects of this invention, treatment of a cancer exhibiting an elevated level of Grb10 phosphorylation can lead to relief of the Grb10-mediated feedback inhibition of PI3K. PI3K signaling, however, promotes proliferation and survival, and aberrant PI3K activity is known to contribute to carcinogenesis. The discovery that some mTOR targets identified herein, for example, Grb10, mediate a feedback inhibition of pro-survival and pro-proliferation signaling (e.g. PI3K, Akt, and MAPK signaling) in cancer cells, could explain why some types of cancer exhibiting elevated levels of mTOR activity do not respond favorably to treatment with an mTORC1 inhibitor (e.g., rapamycin) alone. The mTORC1 inhibitor may cause relief of the feedback inhibition which may, in turn result in sustained survival and proliferation, and even an increase or acceleration in disease progression or recurrence, as observed in some clinical trials of cancer treatment with mTOR inhibitors.
Also provided herein are methods of treating a cancer determined to exhibit an elevated level of Grb10 phosphorylation. In some embodiments, the method comprises administering to the subject an mTOR inhibitor and, additionally, an agent that stabilizes Grb10 or that inhibits the degradation of Grb10. In some embodiments, the method comprises effecting an inhibition of Grb10 degradation by administering a compound that inhibits Grb10 degradation. In some embodiments, the compound that inhibits Grb10 degradation is a ubiquitin ligase inhibitor, for example, a ubiquitin E3 ligase inhibitor. In some embodiments, the method comprises administering to the subject an mTOR inhibitor and, additionally, a PI3K inhibitor. PI3K inhibitors are known to those of skill in the art and described in more detail elsewhere herein.

Diagnostic Methods

Classification of Cancer Cells Based on Grb10 Phosphorylation

Some aspects of this invention provide methods for the classification of a neoplastic disease, for example, a cancer, based on the phosphorylation status of Grb10. In some embodiments, a method is provided that includes obtaining a neoplastic cell from a subject diagnosed to have a cancer, determining the level of Grb10 phosphorylation in the cell, and comparing the level of Grb10 phosphorylation to a reference level. In some embodiments, if the level of Grb10 phosphorylation in the cell is higher than the reference level, then the cell is determined to exhibit an elevated level of mTOR kinase activity. In some embodiments, the cell is determined to exhibit an elevated level of mTORC1 kinase activity based on an elevated level of Grb10 phosphorylation.
Methods of determining a level of protein phosphorylation, for example, of Grb10 phosphorylation, are well known to those of skill in the art. Exemplary methods as well as materials useful for such methods are described, for example, in Marjo de Graauw (Editor), Phospho-Proteomics: Methods and Protocols (Methods in Molecular Biology), Humana Press; 1 edition (2009), ISBN-10: 1603278338. Phosphosensitive binding agents, for example, phosphosensitive antibodies, are provided herein, are commercially available, or can be obtained by those of skill in the art with no more than routine experimentation.
Such methods include, for example, detection of phosphorylated protein with a phosphosensitive antibody and comparison of the amount of phosphorylated protein detected to the total amount of the protein in a sample. Other methods for quantitative or semi-quantitative detection of phosphorylated protein will be apparent to those of skill in the art and the invention is not limited in this respect.
In some embodiments, a method is provided that includes selecting a method of treatment of a neoplastic disease, for example, a cancer based on a cell from a subject having the disease exhibiting an elevated level of Grb10 phosphorylation. In some embodiments, an elevated level of Grb10 phosphorylation is indicative of an elevated level of rapamycin-sensitive mTORC1 kinase activity in the cell. In some embodiments, a method of treatment is chosen based on the cell exhibiting an elevated level of Grb10 phosphorylation that includes administration of an mTORC1 inhibitor, for example, a rapamycin or a rapamycin analog.
Classification of Cancer Cells Known to Exhibit Elevated Levels of mTOR Activity
Some aspects of this invention relate to the discovery that the mTOR target Grb10 is an inhibitor of the mTOR upstream regulators, PI3K, Akt, and MAPK, and, thus, can be characterized as a tumor suppressor gene and an mTOR feedback inhibitor. Some aspects of this invention provide a method for selecting a treatment of a cancer known to exhibit an elevated mTOR kinase activity based on the expression level of the mTORC1 target Grb10. In some embodiments, the method comprises obtaining a cancer cell from a subject diagnosed to have a cancer exhibiting an elevated level of mTOR activity, determining the level of Grb10 expression in the cell, and comparing the level of Grb10 expression to a reference level. In some embodiments, if the level of Grb10 expression in the cancer cell is higher than the reference level, then the cell is determined to exhibit a high likelihood of expressing an elevated level of PI3K, Akt, and/or MAPK activity upon being contacted with an mTORC1 inhibitor. In some embodiments, elevated expression of Grb10 supports the conclusion that upon treatment of the cell or a population of cells of the same type, mTORC1-dependent phosphorylation of Grb10 will be decreased or abolished, resulting in rapid ubiquitination and degradation of Grb10 protein, and, subsequently, in a relief of the Grb10-mediated feedback inhibition of PI3K, as described herein. This relief of feedback inhibition may, in some embodiments, result in elevated PI3K signaling. In some embodiments, elevated PI3K in a cancer cell is an undesirable response to treatment of a subject because it can lead to increased cell proliferation and/or survival, thus offsetting or even outweighing the beneficial effect of the administered mTORC1 inhibitor.
Measuring the expression level of an mTOR target protein can be achieved by using methods well known to those of skill in the art, including, but not limited to, protein expression assays, for example, immunostaining methods (e.g., western blot, protein microarray, immunohistochemistry, phosphoproteomic assays using phosphosensitive binding agents), ELISA, transcript expression assays, for example, RT-PCR, massive parallel sequencing assays, microarray assays, northern blot, or in situ hybridization. Other suitable methods will be apparent to those of skill in the art and the invention is not limited in this respect. See, for example, Sambrook et al., Molecular Cloning, second edition, Cold Spring Harbor Laboratory, Plainview, N.Y.; (1989); Ausubel et al., Current Protocols in Molecular Biology, Current Protocols (1989), and DNA Cloning: A Practical Approach, Volumes I and II (ed. D. N. Glover) IREL Press, Oxford, (1985); and Current Protocols in Molecular Biology, Wiley Publishing, accessible at www.currentprotocols.com; each of which is incorporated herein by reference.
In some embodiments, after a cancer cell is classified, a method of treatment of the subject from which the cell was obtained is selected based on the cancer cell exhibiting a high likelihood of expressing an elevated level of PI3K, Akt, and/or MAPK activity upon being contacted with an mTORC1 inhibitor. In some embodiments, the method of treatment comprises administering an effective amount of an mTOR kinase inhibitor and, additionally, an effective amount of an IGF1R, EGFR, PI3K, Akt, MEK, or RSK inhibitor, or a combination thereof. In some embodiments, the method of treatment comprises administering an effective amount of an mTOR kinase inhibitor and, additionally, of a compound stabilizing Grb10 to the subject. In some embodiments, the mTOR kinase inhibitor is an mTORC1 inhibitor, for example, rapamycin or a rapamycin analog. In some embodiments, the selected method of treatment is communicated to the subject, to a physician or other health care professional treating the subject. In some embodiments, the selected method of treatment is carried out.
Determining and Classifying mTOR Activity in a Cell Based on Phosphoproteomics
Some aspects of this invention provide methods for determining the level of mTOR activity in a cell based on analysis of the phosphorylation state of one or more mTOR targeted phosphorylation sites as provided herein. Some aspects of this invention provide methods to determine a class of mTOR signaling, for example, rapamycin-sensitive mTORC1 signaling, rapamycin-insensitive mTORC1 or mTORC2 signaling, or rapamycin-sensitive and/or rapamycin-insensitive mTOR signaling, in a cell based on an analysis of the phosphorylation state of a phosphorylation site as provided herein.
Methods useful for the determination of the phosphorylation level of a phosphorylation site in a cell are known to those of skill in the art. In some embodiments, such methods comprise obtaining a proteinaceous extract from the cell under conditions that allow for protein phosphorylation to be preserved with high fidelity. In some embodiments, such methods comprise contacting the cell extract with a phosphosensitive binding agent or with a plurality of phosphosensitive binding agents, for example, phosphosensitive antibodies or antibody fragments.
The invention provides phosphosensitive antibodies and antibody fragments to the phosphorylation sites described in the tables and the database. For example, phosphosensitive mTORC1 downstream effector antibodies, including, but not limited to, anti-phospho-NDRG3 (Ser331), anti-phospho-S501/503-Grb10, Anti-CDC2-related Kinase, Arg/Ser-Rich (Ser437), In some embodiments, antibodies provided herein are developed in rabbits. Other phosphosensitive antibodies are described elsewhere herein.
Phosphosensitive antibodies are useful in some embodiments to determine the phosphorylation level of one or more phosphorylation sites disclosed herein. One non-limiting example of such an embodiment is a phosphosensitive protein microarray assay. In some embodiments, the cell extract is contacted with a single phosphosensitive binding agent. In some embodiments, the cell extract is contacted with a plurality of binding agents in parallel. In some such embodiments, the cell is contacted with a microarray comprising a plurality of phosphosensitive binding antibodies or antibody fragments immobilized on a solid surface, for example, a glass surface. Phosphoproteomic assays, arrays, binding agents, and methods for sample preparation and analysis are well known in the art, and exemplary methods are described, for example, in Marjo de Graauw (Editor), Phospho-Proteomics: Methods and Protocols (Methods in Molecular Biology), Humana Press; 1 edition (2009), ISBN-10: 1603278338. The foregoing reference is incorporated herein by reference in its entirety for disclosure of methods and materials useful for the determination of the phosphorylation state of a phosphorylation site on a protein in a cell, tissue, or biological sample.
In some embodiments, the method comprises determining the level of phosphorylation of a phosphorylation site of a protein disclosed in Tables 1 or 2, and comparing the level of phosphorylation to a reference level. If the level of phosphorylation in the cell is higher than the reference level, then the cell is determined to exhibit an elevated level of mTOR kinase activity. If the level of phosphorylation in the cell is equal or lower than the reference level, then the cell is determined to not exhibit an elevated level of mTOR kinase activity. Methods of determining a level of phosphorylation are provided herein, and additional suitable methods will be apparent to those of skill in the art. The invention is not limited in this respect.
Suitable reference levels and methods of determining a reference level will be apparent to those of skill in the art. For example, if the cell is a neoplastic cell or cancer cell obtained from a subject, for example form a biopsy of a solid tumor in the subject, a suitable reference level may be obtained, in some embodiments, from a cell obtained from healthy or non-malignant tissue adjacent to the solid tumor, or a healthy cell of the same tissue of origin as the tumor cells from the same subject or from a different subject. If the cell is obtained from a healthy subject, a suitable reference level may be obtained from a cell of the same cell type obtained from another healthy subject. In some embodiments, a suitable reference level may be an average level or a range of levels observed or expected in cells obtained from healthy subjects that are of the same cell type of the same tissue of origin as the cell in question. In some embodiments, a reference level is a historical level, based on experience or prior experiments, or a level published or otherwise known in the art. Other suitable reference levels are described elsewhere herein and additional reference levels and methods to obtain such levels will be apparent to those of skill in the art.
In some embodiments, a plurality of phosphorylation sites are assayed including a Grb10 phosphorylation site, a pNDRG3 phosphorylation site, a CDK12 phosphorylation site, a FOXK1 phosphorylation site, a ZEB2 phosphorylation site, a LARP I phosphorylation site, an MIB1 phosphorylation site, an IBTK phosphorylation site, and/or a SRPK2 phosphorylation site. In some embodiments, a group of phosphorylation sites of proteins involved in a specific biological pathway are assayed, for example, of proteins known to those of skill in the art to be involved in a biological pathway disclosed in Table 10. In some embodiments, the plurality of phosphorylation sites includes mTOR target sites that were previously known. In some embodiments, the plurality of phosphorylation sites comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 phosphorylation sites disclosed in Table 1. In some embodiments, the plurality of phosphorylation sites comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 phosphorylation sites disclosed in Table 2. In some embodiments, the plurality of phosphorylation sites comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 phosphorylation sites disclosed in Table 3 or in replicate described therein. In some embodiments, the plurality of phosphorylation sites comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 phosphorylation sites disclosed in Table 7 or in replicate described therein. In some embodiments, the plurality of phosphorylation sites comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 phosphorylation sites disclosed in Table 8. In some embodiments, the plurality of phosphorylation sites comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 phosphorylation sites disclosed in Table 11, or in any Rapa or Ku replicate therein
In some embodiments, if a phosphorylation site determined to exhibit a higher level of phosphorylation in a cell in question, for example, a cancer cell obtained from a subject, as compared to a reference level is a class I phosphorylation site, then the cell is determined to exhibit an elevated level of rapamycin-sensitive mTORC1 activity. In some embodiments, if a phosphorylation site determined to exhibit a higher level of phosphorylation in the cell as compared to the reference level is a class II phosphorylation site, then the cell is determined to exhibit an elevated level of rapamycin-insensitive mTORC1 and/or mTORC2 activity. In some embodiments, if a phosphorylation site determined to exhibit a higher level of phosphorylation in the cell as compared to the reference level is a class III phosphorylation site, then the cell is determined to exhibit an elevated level of rapamycin-sensitive mTOR activity, or an elevated level of rapamycin-insensitive mTOR activity, or both. In some embodiments, if a cell is determined to exhibit a higher level of phosphorylation at phosphorylation sites of more than one class, then the cell is determined to exhibit a combination of elevated levels of mTOR signaling of the respective type.
In some embodiments, a method is provided comprising selecting a method of treatment based on the level of phosphorylation of the phosphorylation site(s) assayed. In some embodiments, if the cell is determined to exhibit an elevated level of mTOR kinase activity, then a method of treatment is selected that comprises administering an effective amount of an mTOR kinase inhibitor to the subject. In some embodiments, if the cell is determined to not exhibit an elevated level of mTOR kinase activity, then a method of treatment is selected that does not include administering an mTOR kinase inhibitor. In some embodiments, a method is provided that comprises selecting a method of treatment based on the classification of mTOR target sites that are determined to exhibit an elevated level of phosphorylation in the cell. For example, in some embodiments, if the cell is determined to exhibit an elevated level of rapamycin-sensitive mTORC1 activity, then a method of treatment is selected that comprises administering an effective amount of an inhibitor of rapamycin-sensitive mTORC1 kinase activity to the subject. In some embodiments, if the cell is determined to exhibit an elevated level of rapamycin-insensitive mTORC1 and/or mTORC2 activity, then a method of treatment is selected that comprises administering an effective amount of an inhibitor of rapamycin-insensitive mTORC1 kinase activity to the subject. In some embodiments, if the cell is determined to exhibit an elevated level of rapamycin-sensitive mTOR kinase activity, or an elevated level of rapamycin-insensitive mTOR kinase activity, or both, then a method of treatment is selected that comprises administering an effective amount of an inhibitor of rapamycin-sensitive and rapamycin-insensitive mTOR kinase activity or a combination of an inhibitor of rapamycin-sensitive mTOR kinase activity and an inhibitor of rapamycin-insensitive mTOR kinase activity to the subject.
Classification of a Cancer Based on Grb10 and/or PTEN Expression Levels
Some aspects of this invention relate to the surprising discovery that mutations leading to loss of function of the tumor suppressor Grb10 and mutations leading to loss of function of the tumor suppressor PTEN, are mutually exclusive. Loss of function of either tumor suppressor leads to elevated PI3K activity, which is thought to significantly contribute to carcinogenesis in certain types of cancer. In some embodiments, a method is provided that allows for the classification of a cancer based on Grb10 or PTEN expression levels. In some embodiments, the method comprises determining the level of expression of Grb10 in a cancer cell. In some embodiments, the method comprises determining the level of expression of PTEN in a cancer cell. In some embodiments, the method comprises determining the level of expression of Grb10 and PTEN in a cancer cell.
Expression levels of Grb10 and PTEN can be determined by various methods known to those of skill in the art including, for example methods for determining a level of protein, methods for determining a level of mRNA. Since phosphorylation affects protein stability, for example, stability of Grb10, methods for determining a level of protein phosphorylation may also be useful in assessing protein expression levels. Some such methods for expression analysis are provided herein, and additional methods will be apparent to those of skill in the art. The invention is not limited in this respect.
In some embodiments, the level of expression determined for Grb10 or PTEN is compared to a reference level, for example, to a level observed or expected in a healthy cell of the same cell type or of the same tissue of origin. In some embodiments, if the level of expression of Grb10 or of PTEN in the cell is lower than the reference level, then the cell is likely to exhibit an elevated level of PI3K activity. In some embodiments, if the cell is a neoplastic cell in a subject, a method of treatment with a combination of an mTOR inhibitor and a PI3K inhibitor or with a dual mTOR/PI3K inhibitor is indicated.

Phosphoproteomics Binding Agents and Arrays

Phosphosensitive binding agents that specifically bind mTOR targets are also provided by the present invention. For example, some aspects of this invention provide antibodies and antibody fragments to each of the mTOR-targeted phosphorylation site disclosed in any of the Tables provided herein, for example, in any of Tables 1, 2, 3, 7, 8, or 11, and each such antibody is within the scope of the present invention. For example, some aspects of this invention provide aptamers and/or adnectins that specifically bind mTOR-targeted phosphorylation sites disclosed in Table 1, 2, 3, 7, 8, or 11, and each such aptamer or adnectin is within the scope of the present invention. In some embodiments, a kit is provided that comprises such phosphosensitive binding agents. A kit may also include a buffer, a container, control samples, or instructions.
Some embodiments provide a phosphoproteomics array that includes a plurality of phosphosensitive binding agents, for example, antibodies or antibody fragments, aptamers or adnectins, each of which specifically bind to a phosphorylation site disclosed in Table 1, 2, 3, 7, 8, or 11. In some embodiments the phosphosensitive binding agents are immobilized on a solid substrate, for example, on the surface of a glass slide, a bead, or a microtiter plate. In some embodiments, the plurality of phosphosensitive binding agents comprises binding agents that specifically bind to at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 phosphorylation sites disclosed in Table 1. In some embodiments, the plurality of phosphosensitive binding agents comprises binding agents that specifically bind to at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 phosphorylation sites disclosed in Table 2. In some embodiments, the plurality of phosphosensitive binding agents comprises binding agents that specifically bind to at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 phosphorylation sites disclosed in Table 3. In some embodiments, the plurality of phosphosensitive binding agents comprises binding agents that specifically bind to at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 phosphorylation sites disclosed in Table 7. In some embodiments, the plurality of phosphosensitive binding agents comprises binding agents that specifically bind to at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 phosphorylation sites disclosed in Table 8. In some embodiments, the plurality of phosphosensitive binding agents comprises binding agents that specifically bind to at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, or at least 100 phosphorylation sites disclosed in Table 11.
In some embodiments, an anti-phospho-S501/503-Grb10, anti-mTOR, anti-phospho-mTOR(S2481), anti-Grb10 (human), anti-phospho-Akt (Ser473), anti-Akt, anti-S6K, anti-phospho-S6K (T389), anti-IRS2, anti-PARP, anti-caspase 3, anti-4EBP, anti-4EBP (Ser37/46), anti-phospho-ribosomal protein S6 (Ser235/236), anti-ribosomal protein S6, anti-phospho-ERK1/2, anti-phospho-IRS (Y612), anti-p85 and anti-p110 of PI3K, and/or an anti ERK1/2 antibody or antibody fragment is provided. In some embodiments, phosphosensitive antibodies or antibody fragments are provided for mTOR upstream regulators, including, but not limited to anti-phospho-Akt (Ser473), anti-phospho-ERK1/2, anti-phospho-IRS (Y612), anti-p85, and anti-p110 of PI3K. In some embodiments, phosphosensitive antibodies or antibody fragments are provided for mTOR downstream effectors including, but not limited to anti-phospho-mTOR(S2481), anti-phospho-Akt (Ser473), anti-phospho-S6K (T389), anti-4EBP (Ser37/46), and anti-phospho-ribosomal protein S6 (Ser235/236).
In some embodiments, antibodies against mTORC1 downstream effectors identified for the first time herein are provided, including, anti-phospho-NDRG3 (Ser331), anti-phospho-S5011503-Grb10, and anti-CDC2-related kinase, Arg/Ser-Rich (Ser437).
Phosphosensitive binding agents, their generation and purification, and their use in assays, arrays, and methods for phosphoproteomics analyses of biological samples are well known in the art, and exemplary methods are described, for example, in Marjo de Graauw (Editor), Phospho-Proteomics: Methods and Protocols (Methods in Molecular Biology), Humana Press; 1 edition (2009), ISBN-10: 1603278338, incorporated herein by reference in its entirety. The foregoing reference is incorporated herein by reference in its entirety for disclosure of phosphosensitive binding agents, their generation and purification, and their use in assays, arrays, and methods for phosphoproteomics analyses of biological samples.
Methods of using a microarray comprising a plurality of phosphosensitive binding agents that specifically bind mTOR targets are also provided. In some embodiments, methods of using a phosphoproteomics array to determine mTOR activity in a cell, for example, in a cancer cell, are provided. In some embodiments, the method comprises contacting a proteinaceous sample derived from the cell with the phosphoproteomics array under conditions suitable for a protein expressed in the cell to bind to an antibody or antibody fragment of the array. Such conditions are well known to those of skill in the art and exemplary protocols for phosphoprotein microarrays are described in Marjo de Graauw (Editor), Phospho-Proteomics: Methods and Protocols (Methods in Molecular Biology), Humana Press; 1 edition (2009), ISBN-10: 1603278338, incorporated herein by reference for the disclosure of such methods and conditions. In some embodiments, the method further comprises determining a level of phosphorylated protein bound to a binding agent, for example, an antibody or antibody fragment of the array. In some embodiments, this step includes quantification, absolute or relative to a reference level, of the amount of protein bound to a specific binding agent. In some embodiments, the method includes comparing the level of phosphorylated protein bound to an antibody or antibody fragment of the array to a reference level, wherein if the level phosphorylated protein in the sample derived from the cell is higher than the reference level, then the cell is determined to exhibit an elevated level of mTOR kinase activity.
In some embodiments, methods are provided that include generating a phosphoproteomic profile of the cell. In some embodiments, the profile includes phosphorylation levels of a plurality of proteins, for example, mTOR targets expressed in the cell. In some embodiments, the method further includes comparing the phosphoproteomic profile of the cell with a phosphoproteomic profile of a control cell. In some embodiments, if the phosphoproteomic profile of the cell is similar to that of the control cell, then the cell is determined to exhibit a level of mTOR kinase activity similar to that of the control cell. Methods for comparing phosphoprotein profiles are well known in the art and include, for example, hierarchical clustering methods, supervised and unsupervised learning methods, classification methods, for example class predictor building methods based on phosphoproteomics profiles from cells of known character, and calculation of correlation parameters, such as distance analysis or correlation coefficient calculations. Useful similarity ranges are also known to those of skill in the art and the invention is not limited in this respect. For non-limiting examples of disclosures of such methods and algorithms, see Lim, Y. (2005) Mining the tumor phosphoproteome for cancer markers. Clin Cancer Res 11(9): 3163-3169; Kalume, D. et al. (2003) Tackling the phosphoproteome: tools and strategies Current Opinion in Chemical Biology 7: 64-69; Schmelzle, K. & White, F. (2006) Phosphoproteomic approaches to elucidate cellular signaling networks. Current Opinion in Chemical Biology 17: 406-414; Olsen J V et al. Global, in vivo, and site-specific phosphorylation dynamics in signaling networks. Cell. 2006 Nov. 3; 127(3):635-48; Mumby, M. & Brekken, D. (2005) Phosphoproteomics: new insights into cellular signaling. Genome Biology 6: 230.1-230.7; Zhang et al. J. Proteome Res. vol. 5 pp. 581-8 2006; Hoffert J D, et al. Quantitative phosphoproteomics of vasopressin-sensitive renal cells: regulation of aquaporin-2 phosphorylation at two sites. Proc Natl Acad Sci USA. 2006 May 2; 103(18):7159-64; Johnson, S & Hunter, T. (2004) Phosphoproteomics finds its timing. Nature Biotech 22(9): 1093-1094; and Marjo de Graauw (Editor), Phospho-Proteomics: Methods and Protocols (Methods in Molecular Biology), Humana Press; 1 edition (2009), ISBN-10: 1603278338; all incorporated herein by reference in their entirety
In some embodiments, the control cell is a healthy cell. In some embodiments, the control cell is a cell exhibiting normal mTOR kinase activity. In some embodiments, the control cell is a cancer cell. In some embodiments, the control cell is a rapamycin-sensitive cancer cell. In some embodiments, the control cell is a rapamycin insensitive cancer cell.
Identification of mTOR Kinase Inhibitors Based on mTOR Target Analysis
In some aspects, the invention provides methods of identifying an mTOR kinase inhibitors by analyzing the phosphorylation state of an mTOR target as provided herein after contacting a cell or test sample with a candidate agent. In some embodiments, mTOR phosphorylation efficiency directed towards a target disclosed in Table 1, 2, 3, 7, 8, or 11 is measured in the presence of a candidate agent. In some embodiments, the method includes contacting an mTOR kinase molecule with a polypeptide comprising a phosphorylation site disclosed in Table 1, 2, 3, 7, 8, or 11 under conditions suitable for the mTOR kinase to phosphorylate the phosphorylation site in the presence of a candidate agent. In some embodiments, the level of phosphorylation of the phosphorylation site is then determined and compared to a reference level. In some embodiments, if the level obtained in the presence of the candidate agent is lower than the reference level, then the candidate agent is identified as an mTOR kinase inhibitor.
In some embodiments, the phosphorylation site is a Grb10 phosphorylation site, a pNDRG3 phosphorylation site, a CDK12 phosphorylation site, a FOXK1 phosphorylation site, a ZEB2 phosphorylation site, a LARP1 phosphorylation site, an MIB1 phosphorylation site, an IBTK phosphorylation site, and/or a SRPK2 phosphorylation site. In some embodiments, the candidate agent is a polypeptide, an aptamer, an adnectin, or a small molecule. In some embodiments, the reference level is the level of phosphorylation of the phosphorylation site determined in the absence of the candidate agent. In some embodiments, the level determined in the presence of the candidate agent is lower than the reference level, if the level determined in the presence of the candidate agent is less than 75%, less than 50%, less than 40%, less than 30%, less than 25%, less than 20%, less than 10%, less than 5%, less than 2.5%, or less than 1% of the reference level.
In some embodiments, the contacting is performed in vitro. In some embodiments, the contacting is performed in vivo. In some embodiments, the reference level is a level observed or expected in the absence of the candidate agent. In some embodiments, the reference level is a level observed or expected in the absence of any candidate agent. In some embodiments, the reference level is a level observed or expected in the presence of a known agent. In some embodiments, the reference level is a level observed or expected in the presence of a control agent.

Subjects and Cells

In some embodiments, the subject is an animal. In some embodiments, the subject is a domesticated animal. In some embodiments, the subject is a mammal. In some embodiments, the subject is a non-human mammal. In some embodiments, the subject is a non-human primate. In some embodiments, the subject is a mouse, rat, or rabbit. In some embodiments, the subject is a sheep, goat, cattle, pig, horse, dog, or cat. In some embodiments, the subject is a human.
In some embodiments, the subject is a healthy subject. In some embodiments, the subject is a subject having a hyperproliferative disease. In some embodiments, the subject is a subject having a neoplastic disease. In some embodiments, the subject is a subject having a cancer. In some embodiments, the subject is a subject having a cancer characterized and/or diagnosed to exhibit an elevated level of mTOR activity. In some embodiments, the subject is a subject who had a tumor removed.
In some embodiments, the cell is a healthy cell. In some embodiments, the cell is any cell. In some embodiments, the cell is a bacterial cell. In some embodiments, the cell is an animal cell. In some embodiments, the cell is a mammalian cell. In some embodiments, the cell is a human cell. In some embodiments, the cell is a cell of a cell line. In some embodiments, the cell is a transformed or immortalized cell. In some embodiments, the cell is a neoplastic cell. In some embodiments, the cell is a tumor cell. In some embodiments, the cell is a cancer cell. In some embodiments, the cell is obtained from a tumor in a subject, for example, by tumor biopsy. In some embodiments, the cell is a cell obtained from a tumor that has been removed from a subject.
In some embodiments, the cell is a cell known to exhibit an elevated level of mTOR activity. In some embodiments, the cell is a cell carrying a mutation in a gene involved in an mTOR signaling pathway. In some embodiments, the gene involved in an mTOR signaling pathway is a gene involved in IGF signaling, EGFR signaling, GF signaling, PI3K signaling, AKT signaling, MAPK signaling, Ras signaling, Raf signaling, or Rb signaling. In some embodiments, the gene is TSC1/2, a receptor tyrosine kinases (RTK), PI3K, PTEN, Akt, Ras, Raf, MEK, LKB, or NF2.

Kinase Inhibitors

Kinase inhibitors useful for some aspects of this invention include, but are not limited to, mTOR inhibitors, PI3K inhibitors, Akt inhibitors, and MAPK inhibitors. Further, inhibitors useful for some methods provided herein include, for example, inhibitors of mTOR target degradation, for example, inhibitors of Grb10 degradation. In some embodiments, a kinase inhibitor, for example, an mTOR inhibitor, as provided herein, is an allosteric mTOR kinase inhibitor or a catalytic mTOR kinase inhibitor. In some embodiments, the allosteric mTOR kinase inhibitor is rapamycin or a rapamycin analog. In some embodiments, the catalytic mTOR kinase inhibitor is an ATP-competitive mTOR kinase inhibitor. Other allosteric and catalytic mTOR kinase inhibitors are well known to those of skill in the art, and the invention is not limited in this respect.
In some embodiments, an mTOR kinase inhibitor as provided herein is an mTORC1 inhibitor. In some embodiments, an mTOR kinase inhibitor as provided herein is an mTORC1/2 inhibitor. In some embodiments, an mTOR kinase inhibitor as provided herein is rapamycin or a rapamycin analog. In some embodiments, the rapamycin analog is Ridaforolimus, Sirolimus or Everolimus. In some embodiments, an mTOR kinase inhibitor as provided herein is PP242, PP30, AZD8055, OSI-027, WYE354, INK-128, XL388, torin1, rapamycin (sirolimus), FK506 (tacrolimus), CCI779 (temsirolimus), RAD001 (everolimus), AP23573 (deforolimus, ridaforolimus), S-trans,trans-farnesyl thiosalicylic acid (FTS), FKBP38, PX-866, Theophylline, Caffeine, LY303511, PI-103, 2-(morpholin-1-yl)pyrimido[2,1-aplpha]isoquinolin-4-one, or BEZ235 (NVP-BEZ235)
In some embodiments, an mTOR kinase inhibitor as provided herein is a dual PI3K/mTOR kinase inhibitor. In some embodiments, the dual PI3K/mTOR kinase inhibitor is NVP-BEZ235, BGT-226, XL-765, GSK2126458 or SF1126. Some of the mTOR kinase inhibitors listed immediately above are in various stages of clinical trials. Additional mTOR inhibitors will be apparent to the skilled artisan as they are well known in the art, and it should be appreciated that the invention is not limited in this respect.
In some embodiments, a PI3K inhibitor is provided that is useful in a therapeutic method provided by aspects of this invention. In some embodiments, a PI3K inhibitor as provided herein is ZSTK474, TGX221, GDC0941, or LY294002, XL147, PX147, BKM120, GSK 615, CAL101, PX-866, Quercetin, Tetrodotoxin citrate, Thioperamide maleate, IC87114, PI-103, BEZ235 (NVP-BEZ235), TGX-115, (−)-Deguelin, NU 7026, Myricetin, Tandutinib, SF1126, XL765, D-87503, D-106669, or GSK615. In some embodiments, a PI3K inhibitor provided herein is a dual PI3K/mTOR kinase inhibitor, for example, NVP-BEZ235, BGT-226, XL-765, GSK2126458 or SF1126. In some embodiments, an Akt inhibitor is provided that is useful in a therapeutic method provided by aspects of this invention. In some embodiments, the Akt inhibitor is perifosine, GSK690693, A443654 or MK2206.
In some embodiments, a compound is provided that inhibits the degradation of an mTOR target, for example, of Grb10, NDRG3, CDK12, FOXK1, ZEB2, LARP1, MIB1, IBTK, and/or SRPK2. In some embodiments, the compound that inhibits the degradation of an mTOR target is a ubiquitin ligase inhibitor. In some embodiments, the ubiquitin ligase inhibitor is an E3 ubiquitin ligase inhibitor.

Compositions

In some embodiments, pharmaceutical compositions are provided that comprise an mTOR kinase inhibitor, a PI3K, Akt, or MAPK inhibitor, and/or an agent that inhibits the degradation of an mTOR target, for example, Grb10, NDRG3, CDK12, FOXK1, ZEB2, LARP1, MIB1, IBTK, and/or SRPK2. Pharmaceutical compositions provided herein preferably are sterile and contain an effective amount of one or more therapeutic agents as described herein for producing the desired response in a unit of weight or volume suitable for administration to a patient. If the desired response is amelioration of a hyperproliferative disease, neoplastic disease, or cancer, then the response can, for example, be measured by determining the proliferation of neoplastic or cancer cells in a subject after treatment by, for example, measuring tumor volume, evaluating regression, relapse, or disease symptoms, or by obtaining a cell sample and perform cell counting, flow cytometry, FACS, and other methods well known in the art to be suitable to determine cell proliferation.
In some embodiments, the pharmaceutical compositions as described herein may contain suitable buffering agents, for example, acetic acid in a salt form, citric acid in a salt form, boric acid in a salt form, and/or phosphoric acid in a salt form. The pharmaceutical compositions also may contain, optionally, suitable preservatives, such as ascorbic acid, benzalkonium chloride, benzyl alcohol, m-cresol, chlorobutanol, parabens, EDTA, EGTA, and/or thimerosal. The pharmaceutical compositions may conveniently be presented in unit dosage form and may be prepared by any of the methods well-known in the art of pharmacy.
In some embodiments, a therapeutic method or a method of formulating a kinase inhibitor into a medicament for therapeutic use may include the step of bringing the active agent, for example, a kinase inhibitor as described herein, into association with a carrier which constitutes one or more accessory ingredients. In general, compositions are prepared by uniformly and intimately bringing the active compound(s) into association with a liquid carrier, a finely divided solid carrier, or both, and then, if necessary, shaping the product.
In some therapeutic embodiments, a composition or pharmaceutic preparation provided herein is administered orally to a subject having a cancer. In some embodiments, compositions as described herein that are suitable for oral administration may be presented as discrete units, such as capsules, tablets, lozenges, each containing a predetermined amount of the active compound. Other examples of compositions include suspensions in aqueous liquids or non-aqueous liquids, such as a syrup, elixir, or an emulsion. Examples of compositions for parenteral administration include, without being limited to, sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Examples of aqueous carriers are water, alcoholic/aqueous solutions, emulsions or suspensions, for example, saline and buffered media. Examples of parenteral vehicles are sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, and lactated Ringer's or fixed oils. Examples for intravenous vehicles are fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose), and the like. Preservatives and other additives may also be present such as, for example, antimicrobials, anti-oxidants, chelating agents, and inert gases, and the like.
In some embodiments, a composition comprising a compound or a combination of compounds, useful in this invention, may further comprise an antioxidant to retard oxidation of one or more component. Additionally, the prevention of the action of microorganisms can be brought about by a preservative such as an antibacterial and antifungal agent, including but not limited to parabens (e.g., methylparabens, propylparabens), chlorobutanol, phenol, sorbic acid, thimerosal or combinations thereof.
The compounds useful in the invention, for example the mTOR, PI3K, Akt or MAPK inhibitors provided herein, may be derivatized in various ways. As used herein “derivatives” of the compounds (e.g., small molecule JAK2 and other kinase inhibitors) include salts (e.g., pharmaceutically acceptable salts), any complexes (e.g., inclusion complexes or clathrates with compounds such as cyclodextrins, or coordination complexes with metal ions such as Mn²+ and Zn²+), esters such as in vivo hydrolysable esters, free acids or bases, polymorphic forms of the compounds, solvates (e.g., hydrates), prodrugs or lipids, coupling partners and protecting groups. By “prodrugs” is meant for example any compound that is converted in vivo into a biologically active compound.
The term “pharmaceutically acceptable salt” in this respect refers to the relatively non-toxic, inorganic or organic acid addition salts of agents of the present invention. These salts can be prepared in situ in the administration vehicle or the dosage form manufacturing process, or by separately reacting a purified agent of the invention with a suitable organic or inorganic acid, and isolating the salt thus formed during subsequent purification. Representative salts include the bromide, chloride, sulfate, bisulfate, phosphate, phosphonate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate, glucoheptonate, lactobionate, laurylsulphonate salts, and the like. See, for example, Berge et al. (1977) J. Pharm. Sci. 66:1-19.
The pharmaceutically acceptable salts of the subject agents include the conventional nontoxic salts or quaternary ammonium salts of the compounds, e.g., from non-toxic organic or inorganic acids. For example, such conventional nontoxic salts include those derived from inorganic acids such as hydrochloride, hydrobromic, sulfuric, sulfamic, phosphoric, nitric, and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicyclic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isothionic, and the like.
As set out herein, certain compounds may contain a basic functional group, such as amino or alkylamino, and are, thus, capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable acids. The term “pharmaceutically acceptable salts” in this respect refers to the relatively non-toxic, inorganic or organic acid addition salts of compounds of the present invention. These salts can be prepared in situ in the administration vehicle or the dosage form manufacturing process, or by separately reacting a purified compound of the invention in its free base form with a suitable organic or inorganic acid, and isolating the salt thus formed during subsequent purification. Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, phosphonate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate, glucoheptonate, lactobionate, and laurylsulphonate salts and the like. See, for example, Berge et al. (1977) J. Pharm. Sci. 66:1-19.
The pharmaceutically acceptable salts of the compounds useful in the present invention include the conventional nontoxic salts or quaternary ammonium salts of the compounds, e.g., from non-toxic organic or inorganic acids. For example, such conventional nontoxic salts include those derived from inorganic acids such as hydrochloride, hydrobromic, sulfuric, sulfamic, phosphoric, nitric, and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicyclic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isothionic, and the like.
In other cases, the compounds may contain one or more acidic functional groups and, thus, are capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable bases. The term “pharmaceutically acceptable salts” in these instances refers to the relatively non-toxic, inorganic and organic base addition salts of compounds of the present invention. These salts can likewise be prepared in situ in the administration vehicle or the dosage form manufacturing process, or by separately reacting the purified compound in its free acid form with a suitable base, such as the hydroxide, carbonate or bicarbonate of a pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable organic primary, secondary or tertiary amine. Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts and the like. Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like. See, for example, Berge et al. (1977) J. Pharm. Sci. 66:1-19.
In embodiments where the composition is in a liquid form, a carrier can be a solvent or dispersion medium comprising but not limited to, water, ethanol, polyol (e.g., glycerol, propylene glycol, liquid polyethylene glycol, etc.), lipids (e.g., triglycerides, vegetable oils, liposomes), and combinations thereof. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin; by the maintenance of the required particle size by dispersion in carriers such as, for example liquid polyol or lipids; by the use of surfactants such as, for example, hydroxypropylcellulose; or combinations thereof such methods. In many cases, it will be advisable to include isotonic agents, such as, for example, sugars, sodium chloride, or combinations thereof.

Therapeutic Methods

Some aspects of this invention provide therapeutic methods, for example, methods for the treatment of subjects having a hyperproliferative disease, a neoplastic disease, or a cancer. In some embodiments, a method of treatment is personalized to a specific subject by taking into account the phosphorylation level of at least one an mTOR target site in the subject. In some embodiments, therapeutic methods disclosed herein include administration of an mTOR inhibitor, for example, of an mTORC1 or an mTORC1/2 inhibitor. In some embodiments, therapeutic methods described herein include administration of an mTOR inhibitor and an additional kinase inhibitor, for example, a PI3K inhibitor, a MAPK inhibitor, MEKIERK inhibitor, or an AKT/PKB inhibitor. In some embodiments, therapeutic methods provided herein include administration of a dual mTOR/PI3K inhibitor, or of a combination of an mTOR inhibitor and a PI3K inhibitor.
In some therapeutic embodiments, a composition disclosed herein, for example, a composition comprising an mTOR inhibitor or a composition comprising an mTOR inhibitor and a PI3K inhibitor is administered to a subject having a cancer in an effective amount. An effective amount, in some embodiments, is an amount sufficient to elicit a desired clinical response in the subject. In some embodiments, the desired response is a slowing or inhibiting of the progression of a disorder, for example, of a malignant neoplastic disorder. In some embodiments, this involves slowing the progression of the disease temporarily, although, in more preferable embodiments, it involves halting the progression of the disease permanently.
In some embodiments, the desired response is a permanent reduction of cancer cell proliferation, for example, to a level comparable to a level found in healthy individuals. In some embodiments, a desired response is the induction of cell death in a cancer cell, in a population of cancer cells, or in all cancer cells in a subject. In some embodiments, the desired response is delaying or preventing the manifestation of clinical symptoms characteristic of the disease or condition.
The effect of administering an mTOR inhibitor, either alone or in combination with an additional compound, for example, a PI3K inhibitor, as provided herein, can be monitored by routine methods well known to those of skill in the related medical arts, for example, by methods involving assessment of cancer cell proliferation.
What constitutes an effective amount will depend on the particular condition being treated, the severity of the condition, the individual patient parameters including age, physical condition, size, and weight, the duration of the treatment, the nature of concurrent therapy (if any), the specific route of administration and like factors within the knowledge and expertise of the health care professional treating the subject. These factors are well known to those of ordinary skill in the art and can be addressed with no more than routine experimentation. It is generally preferred that a maximum dose of the individual components or combinations thereof be used, that is, the highest safe dose according to sound medical judgment. It will be understood by those of ordinary skill in the art, however, that a lower dose or tolerable dose may be used for medical reasons.
In general, an effective amount of a therapeutic agent, for example, an mTOR inhibitor or a combination of an mTOR inhibitor with a PI3K, Akt, or MAPK inhibitor, as provided herein, for the treatment of a cancer exhibiting an elevated level of mTOR kinase activity is a dose that achieves an alleviation of the specific neoplastic disease or disorder being treated, for example, by prevention, inhibition, amelioration, delay, or elimination of a symptom of such a disease or disorder.
Some embodiments provide a method of inducing cell death and/or inhibiting proliferation in a neoplastic cell exhibiting elevated mTOR kinase activity by contacting the cell with an mTOR kinase inhibitor or a combination of an mTOR kinase inhibitor and another kinase inhibitor, for example, a PI3K, Akt, or MAPK inhibitor, chosen based on the determination of the type of mTOR kinase signaling active in the cell. In some embodiments, the neoplastic cell is contacted in vivo by administering a composition disclosed herein to a subject carrying the cell. In some embodiments, the neoplastic cell is contacted ex vivo. In some embodiments, the cell is contacted in vitro.
In some embodiments, a neoplastic cell is contacted in vivo, ex vivo, or in vitro, with an effective amount of an mTOR inhibitor or a combination of an mTOR inhibitor and an additional kinase inhibitor, for example, a PI3K, Akt, or MAPK inhibitor, as provided herein. An effective amount, in some embodiments, is an amount sufficient to elicit a desired response in the contacted cell. In some embodiments involving contacting a neoplastic cell exhibiting elevated mTOR activity, for example, because of a mutation in an mTOR upstream pathway, the desired response is a slowing or inhibiting of the proliferation of the cell. In some embodiments, this decreases the proliferation rate and/or cell viability and/or life span, although, in more preferable embodiments, it involves the induction of cell death in the contacted cell or cells.
In some embodiments, the therapeutic methods provided herein further involve the administration of an additional antiproliferative agent to a cancer cell or to a subject carrying a cancer cell, for example, as part of a malignant tumor. Additional antiproliferative agents useful in the methods described herein are well known in the art and include, but are not limited to chemotherapeutic agents (e.g., cytostatic, and cytotoxic agents). Cytotoxic and cytostatic drugs are drugs that kill malignant cells, or inhibit their proliferation, respectively. Examples of cytotoxic and cytostatic drugs include, for example, alkylating agents, antimetabolites, antitumor antibiotics, vinca alkaloids, taxanes, topoisomerase-I compounds, anthrapyrazoles, and epidophylotoxins. In addition, angiogenesis inhibiting drugs, including, for example, compounds that block growth promoting receptors (e.g., PDGF-R and VEGF-R) such as sunitinib (Sutent®) may be used as additional antiproliferative agents. Non-limiting examples of additional antiproliferative agents include Cytoxan® (Cyclophosphamide), Methotrexate, 5-Fluorouracil (5-FU), Adriamycin® (Doxorubicin), Prednisone, Nolvadex® (Tamoxifen), Taxol® (Paclitaxel), Leucovorin, Oncovin® (Vincristine), Thioplex® (Thiotepa), Arimidex® (Anastrozole), Taxotere® (Docetaxel), Navelbine®, (Vinorelbine), Gemzar® (Gemcitabine), Ifex® (Ifosfamide), Pemetrexed, Topotecan, Melphalan (L-Pam®), Cisplatin (Cisplatinum®, Platinol®), Carboplatin (Paraplatin®), Carmustine (BCNU; BiCNU®), Methotrexate, Edatrexate, Mitomycin C (Mutamycin®), Mitoxantrone (Novantrone®), Vincristine (Oncovin®), Vinblastine (Velban®), Vinorelbine (Navelbine®), Fenretinide, Topotecan, Irinotecan, 9-amino-camptothecin (9-AC); Biantrazole, Losoxantrone, Etoposide, and Teniposide.
Administration schedules, formulations, dosages, and administration routes of antiproliferative agents and compositions are well known to those in of skill in the art. Exemplary administration routes, schedules, and dosages of commonly used chemotherapeutic drugs are described in Perry, The Chemotherapy Source Book, 4^thEdition, Lippinkott Williams & Wilkins, 2008, incorporated herein by reference. Such administration schedules may comprise the administration of a single antiproliferative drug or the administration of a combination of such drugs, for example, one of the following, commonly administered combinations: CMF (cyclophosphamide, methotrexate, and 5-fluorouracil); classic CMF (oral cyclophosphamide plus methotrexate and 5-fluorouracil); CAF or FAC (cyclophosphamide, Adriamycin® (doxorubicin), and 5-fluorouracil); AC (Adriamycin® and cyclophosphamide); ACT (Adriamycin® plus cyclophosphamide and tamoxifen); AC taxol (Adriamycin® plus cyclophosphamide and paclitaxel (Taxol®); FACT (5-fluorouracil plus Adriamycin®, cyclophosphamide, and tamoxifen); A-CMF or Adria/CMF (4 cycles of Adriamycin® followed by 8 cycles of CMF); CMFP (CMF plus prednisone); CMFVP (CMF plus vincristine and prednisone); CAFMV (CAF plus methotrexate and vincristine); CMFVATN (CMF plus vincristine, Adriamycin®, thiotepa, and tamoxifen); MF (methotrexate plus 5-fluorouracil and leucovorin). The administration of such combinations of antiproliferative drugs and agents in addition to the administration of an mTOR kinase inhibitor with or without an additional kinase inhibitor, for example, a PI3K, Akt, or MAPK inhibitor, is also envisioned to be embraced by some embodiments.
The therapeutic inhibitors and compositions can be administered in a single dose comprising an effective amount of the individual agents. Multiple doses of the compounds of the invention are also contemplated. When a plurality of inhibitors are used together, they may be administered individually or sequentially, either in a single medicament or in separate units to provide therapeutic doses of the individual compounds. Many mTOR inhibitors, PI3K inhibitors, Akt inhibitors and MAPK inhibitors described herein are in clinical studies or even in clinical use. Therapeutic doses of such compounds are, accordingly, well known in the field of medicine. Dosages of compounds in clinical use are described in references such as Remington's Pharmaceutical Sciences, 18th ed., 1990; as well as many other medical references relied upon by the medical profession as guidance for the treatment of proliferation disorders.

Administration Routes and Methods

A variety of administration routes are available for the kinase inhibitors and other therapeutic agents described herein. The particular mode selected will depend, of course, upon the particular compound selected, the particular condition being treated and the dosage required for therapeutic efficacy. The methods of this invention may be practiced using any mode of administration that is medically acceptable, meaning any mode that produces effective levels of compounds without causing clinically unacceptable adverse effects. Examples of modes of administration are parenteral routes. The term “parenteral” includes subcutaneous, intravenous, intramuscular, intraperitoneal, and intrasternal injection, or infusion techniques. Other routes include, but are not limited to, oral, nasal, dermal, sublingual, and local.
The formulations of the invention are administered in pharmaceutically acceptable solutions, which may routinely contain pharmaceutically acceptable concentrations of salts, buffering agents, preservatives, compatible carriers, adjuvants, and optionally other therapeutic ingredients.
According to the methods provided by aspects of the invention, the compounds described herein may be administered in a pharmaceutical composition. In some embodiments, a pharmaceutical composition comprises a compound provided by aspects of the invention and a pharmaceutically acceptable carrier. As used herein, a pharmaceutically acceptable carrier refers to a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredients.
Pharmaceutically acceptable carriers include diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials which are well known in the art. Such preparations may routinely contain salt, buffering agents, preservatives, compatible carriers, and optionally other therapeutic agents. When used in medicine, the salts should be pharmaceutically acceptable, but non-pharmaceutically acceptable salts may conveniently be used to prepare pharmaceutically acceptable salts thereof and are not excluded from the scope of the invention. Such pharmacologically and pharmaceutically acceptable salts include, but are not limited to, those prepared from the following acids: hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, maleic, acetic, salicylic, citric, formic, malonic, succinic, and the like. Also, pharmaceutically acceptable salts can be prepared as alkaline metal or alkaline earth salts, such as sodium, potassium or calcium salts.
The compounds used in the invention may be formulated into preparations in solid, semi-solid, liquid or gaseous forms such as tablets, capsules, powders, granules, ointments, solutions, depositories, inhalants and injections, and usual ways for oral, parenteral or surgical administration. Some aspects of the invention also embrace pharmaceutical compositions which are formulated for local administration, such as by implants.
Compositions suitable for oral administration may be presented as discrete units, such as capsules, tablets, lozenges, each containing a predetermined amount of the active compound. Other compositions include suspensions in aqueous liquids or non-aqueous liquids such as a syrup, elixir or an emulsion. When the compounds described herein are used therapeutically, in certain embodiments a desirable route of administration may be by pulmonary aerosol.
In some embodiments, a compound provided by some aspects of the invention may be administered directly to a tissue. Direct tissue administration may be achieved by direct injection. A compound may be administered once or alternatively may be administered in a plurality of administrations. If administered multiple times, a compound may be administered via different routes. For example, the first (or the first few) administrations may be made directly into the affected tissue while later administrations may be systemic.
For oral administration, the compounds can be formulated readily by combining the active compounds with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a subject to be treated. Pharmaceutical preparations for oral use can be obtained as solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate. Optionally the oral formulations may also be formulated in saline or buffers for neutralizing internal acid conditions or may be administered without any carriers.
Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.
Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. Microspheres formulated for oral administration may also be used. Such microspheres have been well defined in the art. All formulations for oral administration should be in dosages suitable for such administration.
For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.
For administration by inhalation, the compounds for use according to the present invention may be conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of, e.g., gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.
The compounds, when it is desirable to deliver them systemically, may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and/or dispersing agents.
Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose), and the like. Preservatives and other additives may also be present such as, for example, antimicrobials, anti-oxidants, chelating agents, and inert gases and the like. Lower doses will result from other forms of administration, such as intravenous administration. In the event that a response in a subject is insufficient at the initial doses applied, higher doses (or effectively higher doses by a different, more localized delivery route) may be employed to the extent that patient tolerance permits. Multiple doses per day are contemplated to achieve appropriate systemic levels of compounds.
Other delivery systems can include time-release, delayed release or sustained release delivery systems. Such systems can avoid repeated administrations of the compound, increasing convenience to the subject and the physician. Many types of release delivery systems are available and known to those of ordinary skill in the art. They include polymer base systems such as poly(lactide-glycolide), copolyoxalates, polycaprolactones, polyesteramides, polyorthoesters, polyhydroxybutyric acid, and polyanhydrides. Microcapsules of the foregoing polymers containing drugs are described in, for example, U.S. Pat. No. 5,075,109. Delivery systems also include non-polymer systems that are: lipids including sterols such as cholesterol, cholesterol esters and fatty acids or neutral fats such as mono- di- and tri-glycerides; hydrogel release systems; silastic systems; peptide based systems; wax coatings; compressed tablets using conventional binders and excipients; partially fused implants; and the like. Specific examples include, but are not limited to: (a) erosional systems in which the platelet reducing agent is contained in a form within a matrix such as those described in U.S. Pat. Nos. 4,452,775, 4,675,189, and 5,736,152 and (b) diffusional systems in which an active component permeates at a controlled rate from a polymer such as described in U.S. Pat. Nos. 3,854,480, 5,133,974 and 5,407,686. In addition, pump-based hardware delivery systems can be used, some of which are adapted for implantation.
Therapeutic formulations useful in the invention may be prepared for storage by mixing a kinase inhibitor having the desired degree of purity with optional pharmaceutically acceptable carriers, excipients or stabilizers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the form of lyophilized formulations or aqueous solutions. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes); and/or non-ionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG).

EXAMPLES

Two sets of large-scale, quantitative phospho-proteomics experiments were performed to fully define how mTOR-containing complexes signal to downstream effectors (FIG. 1A). The first SILAC experiment (rapamycin screen) was performed using growth factor-deprived TSC2−/− mouse embryonic fibroblasts (MEFs). Deletion of the TSC2 tumor suppressor gene decouples mTORC1 from many upstream inputs, leading to constitutive hyper-activation of mTORC1 signaling (7). mTORC1 is still potently and specifically inhibited by rapamycin in these cells, which provides a sensitized genetic background for the study of mTORC1 signaling in the absence of other mitogen-regulated phosphorylation cascades.
Global Quantitative Phosphoproteomics Defines the Signaling Networks Downstream of both mTORC1 and mTORC2
Two populations of TSC2−/− cells were grown in parallel, one in conventional media (“light”) and the other in media containing [¹³C₆ ¹⁵N₂]lysine and [¹³C₆ ¹⁵N₄]arginine (“heavy”) (FIG. 1A and Table 5). As shown in FIG. 5A, phosphopeptides were successfully enriched using a two-step SCX-IMAC procedure with the phosphopeptides representing 67.8% of the peptides identified. We performed two biological replicates of this experimental design with cross-labeling (swapping the labeled state of the rapamycin-treated cells, Table 5), from which a total of 14,635 (FDR=0.25%) and 32,500 (FDR=0.28%) phosphopeptides were identified, respectively. These corresponded to identification of at least 4,484 and 6,832 unique phosphorylation sites on 1,615 and 1,866 proteins, respectively. The quantitation was highly accurate (Table 5) with 93.7% of the phosphoproteome changing less than 50% in abundance between the rapamycin-treated (light) and control (heavy) cells (second biological replicate experiment, Median Log₂(H/L)=−0.02 and standard deviation Log₂(H/L)=0.53, FIG. 1A). Gene names and sequences of the phosphopeptides whose intensities decrease after rapamycin treatment (Rapamycin screen) are identified in Table 7. Note that the Rapamycin screen includes two biological replicates. For the first replicate (Table 7 Rapa Replicate 1), the light cells were controls whereas the heavy cells were treated with rapamycin. For the second replicate experiment (Table 7 Rapa Replicate 2), the light cells were treated with rapamycin and heavy cells were controls.
Based on the tight distribution of the quantified phosphopeptides, we considered phosphorylated peptides whose relative abundance decreased ≧2-fold as regulated in a rapamycin-sensitive fashion. Using this criteria, several hundred peptides (corresponding to 148 and 85 proteins respectively, in the two biological replicates, Table 11) were determined to contain rapamycin-sensitive phosphorylation sites. Table 11 identifies proteins with downregulated phosphorylation identified in the rapamycin and Ku-0063794 screen. Note that there are two biological replicate experiments for the rapamycin screen (“Table 11 Rapa Replicate 1,” and “Table 11 Rapa Replicate 2,” respectively) and one Ku-0063749 replicate (“Table 11 Ku”). Treatment for the light and heavy cells in each experiment is described in FIG. 1C. We observed a similar distribution for the phosphopeptides in the first biological replicate experiment and found 85 proteins (Table 11) that carried rapamycin-sensitive phosphorylation sites. In addition, there was a substantial overlap in the down-regulated proteins between replicates and 40 of these proteins were found in both experiments (FIG. 5B).
For the Ku-screen, wild-type MEFs were grown in light and heavy SILAC media and starved overnight for serum. The light cells were treated with 20 nM rapamycin for 2 hrs, while the heavy cells were treated in parallel with a combination of 20 nM rapamycin and 2 μM of Ku-0063794 for 2 hrs. Both the light and heavy cell were subsequently stimulated with insulin for 15 minutes and samples were pooled and analyzed by quantitative mass spectrometry. Ku-0063794 is a recently identified compound that competitively inhibits mTOR kinase activity by occupying the ATP-binding pocket (FIG. 5C) (4, 11). For the Ku-screen, a total of 34,642 phosphopeptides were identified with a false discovery rate of 0.28%, which corresponded to identification of 6,220 unique phosphorylation sites on 1,867 proteins (Table 5). Gene names and sequences of the phosphopeptides whose intensities decrease after Ku-0063794 treatment are identified in Table 8. Note that the light cells were treated with rapamycin and serve as controls whereas the heavy cells were treated with a combination of rapamycin and Ku-0063794. A very tight distribution of phosphopeptides was observed (Median Log₂(H/L)=−0.03 and standard deviation Log₂(H/L)=0.34) and phosphorylated peptides whose relative abundance decreased ≧2-fold were considered regulated by Ku-sensitive, but rapamycin-insensitive mTOR signaling (FIG. 1A). Using these criteria, 100 proteins were determined to contain downregulated phosphorylation after Ku-0063794 treatment (Table 11). The identified phopho-proteins included many known downstream targets of mTORC2 (Table 6), including GSK3β, Braf, Akt1s1 (Akt substrate) and NDRG1 (SGK substrate). Combining the data from both the Rapa-screen and the Ku-screen, we identified a total of 81,777 phosphopeptides, and more than 11,271 unique phosphorylations sites on 2,778 proteins, with 9,694 of the sites confidently localized (A score ≧13) (Table 5).
Phosphorylation Specificity of the Downstream Effectors in the mTORC1 and mTORC2 Signaling Networks
Table 9 describes the classification of mTOR targets identified in the rapamycin and Ku-0063794 screens. Class I includes downstream effectors of rapamycin-sensitive mTORC1. Class II includes downstream effectors of rapamycin-insensitive mTORC1 or mTORC2. Class II includes the proteins downstream of both mTORC1 and mTORC2. Names of the genes and the sequences of the phosphopeptides are shown.
Class I sites represent phosphorylation events that are mediated by rapamycin-sensitive mTORC1/S6K signaling, (e.g. phospho-rpS6 S235/S236 phosphorylation).
Class II sites represent phosphorylation events mediated by rapamycin-insensitive mTORC1 or mTORC2 function. For example, 4EBP1 T36/T45 phosphorylation, previously characterized as a rapamycin insensitive mTORC1 substrate (12), only decreased slightly (20%) after rapamycin treatment, while phosphorylation at these sites decreased dramatically (7.2-fold) in the Ku-0063794 screen (FIGS. 1C, 5C and 6A). As another example, NDRG1 was also identified to contain Class II phosphorylation sites at S330/S333 (FIGS. 1C and 6B) and was recently shown to be a substrate of SGK (8), whose activation is under the control of mTORC2.
Class III represents phosphorylation sites that are both the rapamycin-sensitive and Ku-sensitive mTOR substrates. For example, GSK3β S9 phosphorylation is down-regulated by approximately 3.3-fold and 2.2-fold in the rapamycin and Ku-0063794 screens, respectively (FIGS. 1C and 6C), consistent with the previous observation that it can be a substrate of both Akt and S6K (9). As another example for the Class III phosphorylation, the abundance of mTOR autophosphorylation at S2478/S2481 decreased 3.6-fold in the rapamycin screen (FIG. 7A). Surprisingly, it was previously shown that S2481 is a conserved, rapamycin-insensitive, autophosphorylation site of mTOR (FIG. 7B) (20). Immunoblot analysis confirmed that site was indeed rapamycin-sensitive (FIGS. 5C and 7C). Intriguingly, acute rapamycin treatment only led to partial dephosphorylation of this site. In contrast, mTOR S2481 phosphorylation was completely abolished as a result of mTOR kinase inhibitor treatment (FIGS. 5C and 7D). Taken together, the data demonstrate that mTOR S2481 phosphorylation is regulated in both a rapamycin sensitive- and insensitive-manner.
There were several surprising examples of phosphopeptides that are insensitive to both rapamycin and Ku-0063794 as identified by the two phosphoproteomics screens. The intensity of a T70 singly phosphorylated 4EBP 1 peptide did not change in either the rapamycin or the Ku-0063794 screen (FIGS. 1C and 6A), indicating the 4EBP T70 is not phosphorylated by mTOR in this context. Indeed, several reports suggested that this site might be targeted by ERK2, another proline-directed kinase (21). In addition, we found that the 4EBP-1 S85/S 100 phosphorylation was also mTOR-independent (FIG. 1C).
Linking Phosphorylation Events to the Biological Processes Regulated by mTORC1 and mTORC2
To assess the predicated effects of mTOR inhibition in cells as a result of either rapamycin or Ku-0063794 treatment, we mapped the downregulated proteins in the two screens to Gene Ontology (GO) terms with respect to Biological Process (BP) and also performed gene functional classification analysis of these proteins. Table 10 describes the results of GO analysis of the hits identified in the rapamycin and Ku-0063794 screens. Pathways that the rapamycin-sensitive and Ku-0063794-sensitive hits overrepresented are shown in “Table 10 Rapa pathways” and “Table 10 Ku pathways”, respectively. Also shown is the biological process that the rapamycin-sensitive and Ku-0063794-sensitive hits overrepresented (Table 10 Rapa BP GO, and Table 10 Ku BP GO, respectively). Interestingly, for the rapamycin screen, the top enriched BPs included negative regulation of macromolecule metabolic process (P=4.2×10⁻⁷), negative regulation of cellular biosynthetic process (P=4.2×10⁻⁶), response to insulin stimulus (P=2.6×10⁻⁴), negative regulation of transcription (P=2.7×10⁻⁴) and vesicle-mediated transport (P=6.0×10⁻⁴). By grouping the hits based on functional similarities, we also found that the downregulated proteins in the rapamycin screen were enriched for kinases (P=2.1×10⁻²⁰), WD40-repeat containing proteins (P=8.9×10⁻¹²), proteins involved in transcription regulation (P=3.1×10⁻⁹) and proteins involved in RNA processing (P=3.3×10⁻⁵). The identification of many kinases in the downregulated proteins provides interesting points for potential signal integration and crosstalk. One example is the Ser/Thr kinase, unc-51-like kinase 1 (ULK1). Previously published research has demonstrated that ULK1 is positively involved in autophagy response (22). ULK1 has also been shown to interact with mTORC1 through binding to raptor (23). We identified three phosphorylation sites on ULK1 (S747/S757/T763) that showed a 4-fold downregulation after rapamycin treatment (Table 1), suggesting that ULK1 is likely a bona fide mTORC1 substrate. As mTORC1 is known to antagonize macroautophagy, it is tempting to speculate that mTORC1 negative regulates ULK1 either directly or indirectly via inhibitory phosphorylation events. Interestingly, in mammalian cells, knockdown of the related kinase ULK2 had no effect on the autophagic response (22). Interestingly, all three of the rapamycin-sensitive ULK1 phosphorylation sites are proline-directed (SP or TP), whereas two of the three comparable sites in ULK2 possess a C-terminal alanine, making it unlikely that these residues are similarly phosphorylated by mTORC1 (FIG. 7E).
For the downregulated proteins identified in the Ku-0063794 screen, the top enriched BPs included lamellipodium assembly (P=0.003), protein amino acid phosphorylation (P=0.007), the insulin receptor signaling pathway (P=0.01), actin filament-based process (P=0.015) and nucleocytoplasmic transport (P=0.015). Gene functional classification analysis revealed that the downregulated proteins were enriched for ATP-binding proteins (P=2.4×10⁻¹¹) and zinc-finger domain containing proteins (P=2.1×10⁻⁴). Intriguingly, it has been suggested that mTORC2 promotes organization of the actin cytoskeleton upon growth factor stimulation through a mechanism that is poorly defined. The identification of a total of 10 proteins (CCDC88A, ARHGEF17, NCK1, myo9b, Mtap1b, Epb4.113, vel, Spnb2, FLNC, Npm1) in the Ku-0063794 screen that are related to cytoskeleton regulation provides an opportunity for the discovery of novel points of regulation. For example, we identified a peptide from filamin-C (FLNC) in the Ku-0063794 screen that was singly phosphorylated at S2234 (FIG. 2). S2234 is localized in an AGC kinase substrate motif (RERLGS*F, SEQ ID NO: 22) and has previously been shown to be phosphorylated by Akt in vitro (24). We found S2234 could be categorized as a Class II phosphorylation site (rapamycin-insensitive and Ku0063794-sensitive, FIG. 2 and Table 1), suggesting that it is not an S6K substrate but rather the substrate of Akt or SGK. FLNC crosslinks actin filaments into a three-dimensional network and is involved in assembling signaling complexes near the cell membrane (25). Interestingly, filamin-A (FLNA) has been shown to be phosphorylated at a similar site (S2152, RRRAPS*V, SEQ ID NO: 23) by RSK and Pak protein kinases which regulates FLNA function in cell migration (26). It will clearly be of great interest to investigate whether mTORC2 regulates actin assembly and cell migration through Akt- or SGK-mediated phosphorylation of FLNC and whether this phosphorylation coordinates regulation of these processes with Ras-, Cdc42- and Rac-mediated phosphorylation of FLNA.
The analysis identified 4,484 and 6,832 unique phosphorylation sites on 1,615 and 1,866 proteins from two biological replicate experiments, respectively, and achieved a high level of accuracy in quantitation (Table 5). Several hundred peptides corresponding to 148 and 85 proteins in the two replicates (Table 11) were determined to contain rapamycin-sensitive phosphorylation sites (defined as phosphorylated peptides whose relative abundance decreased ≧2-fold in response to rapamycin treatment). There was a substantial overlap in the downregulated proteins between the replicates, as 40 proteins were found in both experiments (FIG. 5B). Supporting the validity of the approach, many known effectors of the mTORC1 signaling pathway were identified in the downregulated population (Table 6), including p70S6K, 4EBP1/2, Akt1s1 (PRAS40), rpS6, eIF4B, eIF4G1 and GSK3β. A representative identification of the known rapamycin-sensitive phosphorylation sites on ribosomal protein S6 is shown in FIG. 1B.
Rapamycin is an allosteric inhibitor that only partially inhibits mTORC1 signaling and has no effect on the activity of mTORC2 under short-term treatment conditions (3). In contrast, newly discovered ATP-competitive mTOR inhibitors block the activity of both mTORC1 and mTORC2 (2). To identify rapamycin-insensitive mTORC1, and mTORC2 substrates, we used the mTOR kinase inhibitor Ku-0063794 and performed a second SILAC experiment (Ku-0063794 screen) (FIG. 1A). The light cells were treated with 20 nM rapamycin for 2 hrs, while the heavy cells were treated in parallel with a combination of 20 nM rapamycin and 2 μM of Ku-0063794 for 2 hrs. Both the light and heavy cells were subsequently stimulated with insulin for 15 minutes and samples were pooled and analyzed by quantitative mass spectrometry. By creating a rapamycin-inhibited background in both the light and heavy cells, the combination of insulin stimulation and treatment with the mTOR kinase inhibitor leads to the identification of proteins specficially phosphorylated by rapamycin-insensitive mTOR signaling and kinases activated downstream of mTORC2 such as Akt and SGK.
In this experiment, one hundred proteins were determined to contain down-regulated phosphorylation after Ku-0063794 treatment (Table 11). The identified phospho-proteins included many known downstream targets of mTORC2 (Table 6), including GSK313, Braf, Akt1s1 (Akt substrates) and NDRG1 (SGK substrate).
To identify the specific downstream effectors of the two mTOR complexes, we compared the fold-change in phospho-peptides identified in both the rapamycin and Ku-0063794 screens (FIG. 1C). As expected, the majority of phospho-peptides in the cell are not affected by either rapamycin or Ku-0063794 treatment and show a heavy:light ratio close to 1:1 in both screens. However, there are clearly phosphopeptides that are mediated by mTORC1 and/or mTORC2 signaling, which can further be divided into three categories. Class I sites include rapamycin-sensitive events that are not further decreased by Ku-0063794 treatment, and represent canonical mTORC1 downstream effectors, such as rpS6 S235/236 phosphorylation (FIG. 1C). Class II sites include Ku-0063794-sensitive events that are not affected by rapamycin treatment and represent either rapamycin-resistant mTORC1 downstream effectors, such as 4EBP1 T36/T45 (FIGS. 1C, 5C and 6A), or sites downstream of the mTORC2 signaling, such as the recently described SGK-mediated NDRG1 S330/S333 phosphorylation events (FIGS. 1C and 6B) (8). Finally, Class III sites are sensitive to both compounds. For example, GSK3β S9 phosphorylation is downregulated in both the rapamycin and Ku-0063794 screens (FIGS. 1C and 6C), consistent with the previous observation that GSK313 can be a substrate of both Akt and S6K (9). In addition, we identified mTOR autophosphorylation at S2481 as a Class III site that is regulated in both a rapamycin sensitive- and insensitive-manner (FIG. 7) (these target classes are discussed in more detail elsewhere herein).
We performed pathway analysis using DAVID (10), to determine the signaling networks that are statistically overrepresented by the hits in the rapamycin and Ku-0063794 screens (FIGS. 1D and 5D). Not surprisingly, proteins with downregulated phosphorylation in both screens were highly enriched for the canonical mTOR pathway (P=1.6×10⁻⁶, ranked first, for the rapamycin screen and P=0.0014, ranked second, for the Ku-0063794 screen).
Interestingly, one of the enriched GO classes in the rapamycin screen is the transmembrane receptor protein tyrosine kinase (RTK) signaling pathway (P=0.01), suggesting that mTORC1 might inhibit proteins in the upstream PI3K and MAPK pathways through modulating the activities of these RTKs. In particular, we observed that phosphorylation of two sites on the growth factor receptor-bound protein 10 (Grb10, S501/S503, ⁴⁹⁶MNILSS*QS*PLHPSTLNAVIHR⁵¹⁶(SEQ ID NO: 24), mass error=2.32 ppm, Xcorr=4.39) was strongly inhibited by rapamycin (FIGS. 2A and 8A). The level of phosphorylation decreased by more than 27-fold after a 2 h rapamycin treatment (Table 6). The intensity of a triply phosphorylated Grb10 peptide (T76/S96/S104, insufficient MS/MS fragment ions to localize T76) also decreased by about five-fold after rapamycin treatment (Table 6). In contrast, phosphorylation of two other Grb10 sites, 5455 and 5458, did not change after a 2-hr rapamycin treatment (FIG. 8B), suggesting that decreased S501/S503 phosphorylation was not the result of a change in Grb10 protein abundance.
Grb10 belongs to the growth factor receptor-bound (Grb) protein family, which contains Grb7, Grb10 and Grb14. Members of this protein family serve as cellular adaptor proteins that bind to activated receptor tyrosine kinases (11). Grb10 has an N-terminal Ras-associating (RA) domain, a PH domain, a C-terminal SH2 domain and a BPS (between PH and SH2) domain, in which the two rapamycin-sensitive phosphorylation sites reside (FIG. 2B). The S501/S503 sites, and their flanking sequences, are highly conserved in vertebrates (FIG. 2B), suggesting that they could be functionally relevant and phosphorylated by a common evolutionarily conserved kinase, such as mTOR.
We developed a phosphospecific antibody (FIGS. 9A and 9B) to further characterize these two Grb10 phosphorylation sites. Treatment of TSC2−/− MEFs with 20 nM rapamycin induced rapid dephosphorylation of Grb10, within 15 min, and remained completely inhibited for the remainder of the time course examined (FIG. 2C). The dephosphorylation kinetics correlated well with that of S6K. We also found that Grb10 phosphorylation at S501/S503 is sensitive to amino acid availability in TSC2−/− MEFs (FIG. 2D).
To determine whether the Grb10 S501/S503 sites can be phosphorylated by other kinases, we treated TSC2−/− cells with staurosporine, a broad-spectrum kinase inhibitor that does not suppress mTOR activity (12). No change in the phosphorylation level of Grb10 S501/S503 phosphorylation was observed in response to staurosporine treatment (FIG. 2E). In contrast, treatment of Ku-0063794 led to a dose-dependent dephosphorylation of Grb10, correlating with the dephosphorylation of mTOR, 4EBP and rpS6. Interestingly, S6K activity was inhibited by staurosporine treatment, as shown by a complete loss of rpS6 phosphorylation, suggesting that Grb10 S501/S503 was directly phosphorylated by mTORC1 rather than by S6K.
We further investigated the effects of growth factor stimulation and rapamycin-mediated inhibition on Grb10 phosphorylation in other cell types. Wild-type MEFs were serum-starved and then stimulated with either insulin or 10% serum, both of which led to a robust increase in Grb10 phosphorylation (FIG. 2F). The insulin- or serum-induced increase in Grb10 phosphorylation was completely blocked by rapamycin pre-treatment. It was previously reported that Grb10 S503 (S476 in human Grb10 isoform 3) was phosphorylated by ERK1/2 in vitro (13). We found that inhibiting MEK by using AZD6244 completely abolished the activities of ERK but had no effect on Grb10 S501/S503 phosphorylation (FIG. 2F), indicating that phosphorylation at these two sites on Grb10 is not mediated by ERK in vivo. We also tested other mTOR catalytic site inhibitors, including LY294002, NVP-BEZ235, torin and pp 242 (14), all of which completely abolished Grb10 S501/S503 phosphorylation (FIG. 2G).
We next examined the in vivo interaction between Grb10 and the components of the mTOR complexes. We co-overexpressed HA-tagged Grb10 with Myc-tagged raptor or rictor in HEK293T cells. Grb10 was found to interact with raptor, but not rictor, suggesting Grb10 is a binding partner of mTORC1, but not mTORC2 (FIG. 3A). To further evaluate the possibility that Grb10 is a substrate of mTORC1, we prepared recombinant Grb10 from bacteria and subjected it to an in vitro kinase assay using recombinant mTOR. As shown in FIG. 3B, Grb10 was robustly phosphorylated by recombinant mTOR at S501/S503.
We found long-term inhibition of mTORC1 by rapamycin led to a significant decrease in Grb10 protein levels (FIG. 3C) while Grb10 mRNA levels showed a modest (approximately 2-fold) decrease, suggesting mTORC1 activity positively regulates Grb10 expression at both the transcriptional and post-translational levels. Similarly, knockdown of the mTORC1 component raptor led to a decreased level of Grb10 protein, correlating with diminished mTORC1 activity, as shown by phospho-4EBP levels (FIG. 3D). In addition, long-term treatment with mTOR kinase inhibitors, including LY-294002, NVP-BEZ235, torin and pp 242 all led to reduced Grb10 protein expression (FIG. 9C). Similar results were obtained in TSC1−/− MEFs (FIG. 9D). In contrast, we found Grb10 was highly overexpressed in TSC2−/− and TSC1−/− MEFs compared with their wild-type counterparts (FIGS. 3E and 9E), correlating with elevated mTORC1 activity in these cells.
To explore whether mTORC1-mediated Grb10 stabilization depends on phosphorylation on S501/S503, we transfected WT-Grb10, Grb10-S501A/S503A (AA) mutant and Grb10-S501D/S503D (DD) mutant into HEK293T cells. Exogenous wild type (WT) and DD mutant Grb10 proteins were expressed at equal levels, while expression of the AA mutant was markedly reduced, suggesting lower protein stability for the unphosphoryated form of Grb10 (FIG. 3F). To confirm this result, we generated TSC2−/− MEFs stably expressing the HA-tagged Grb10-DD mutant and treated these cells with rapamycin. Long-term rapamycin treatment decreased endogenous, wild-type Grb10 levels but had no effect on the DD mutant protein levels (FIG. 3G). To rule out the possibility that this result was an artifact based on protein overexpression, we repeated the experiment on TSC2−/− cells expressing HA-tagged wild-type Grb10, in which rapamycin treatment decreased both the endogenous and ectopically expressed Grb10 (FIG. 9F). These data support a critical role for mTORC1 in stabilizing Grb10 through phosphorylation of the S5011S503 residues.
Grb10 has been suggested to function as a negative regulator of the insulin signaling pathway. In mice with genetically disrupted Grb10 function, overgrowth of both the embryo and placenta was observed, and these mice are approximately 30% larger than normal at birth (15). In addition, enhanced activity of the insulin-stimulated PI3K/Akt pathway was observed in insulin target tissues, including skeletal muscle and adipose tissue (16). We examined the role of mTORC1-mediated Grb10 protein stabilization and accumulation in mTORC1 mediated negative feedback on the PI3K/Akt pathway.
As expected from previous reports (5, 6), both the PI3K/Akt and MAPK pathways were highly suppressed in TSC2−/− cells due to constitutively elevated mTORC1 activity. In contrast, Akt was strongly activated in Grb10 knockdown cells under both serum-starved and insulin/IGF stimulation conditions (FIG. 4A). In addition, Grb10 knockdown also led to ERK hyperactivation upon insulin/IGF stimulation. Conversely, overexpression of Grb10 in HEK293 cells substantially interfered with activation of PI3K, as judged by decreased Akt T308 phosphorylation (FIG. 10A). The observed inhibition was not affected by the phosphorylation status of S501/S503. Previous work demonstrated that Grb10 interferes with insulin-mediated PI3K activation by binding to, and inhibiting, the insulin receptor (11). We confirmed this observation and demonstrated that Grb10 overexpression interferes with insulin receptor-dependent IRS phosphorylation and subsequent PI3K recruitment (FIGS. 10B, 10C and 10D).
The current model of the negative feedback loop from mTORC1 to PI3K involves S6K-mediated phosphorylation and subsequent degradation of IRS 1/2 (6). However, it has also been shown that overexpression of a dominant-negative, kinase-deficient S6K1 failed to completely recapitulate the hyperactivation of Akt induced by overexpression of kinase-dead mTOR, suggesting mTORC1 is directly involved in modulating this feedback inhibition loop (17). Our data clearly show that hyperactivation of mTORC1/S6K promotes feedback loop inhibition of PI3K through a two-prong mechanism: (1) mTORC1/S6K-mediated phosphorylation and degradation of a positive regulator of PI3K signaling, IRS, and (2) mTORC1-mediated phosphorylation and accumulation of a negative regulator of PI3K signaling, Grb10.
Numerous genetic alterations in cancers result in hyperactivation of the mTOR complexes. Based on these observations, rapamycin analogues are currently approved, or being assessed in various clinical trials, as targeted therapeutics against several cancer subtypes. However, with a few exceptions, the current results have been disappointing, as the clinical outcome of rapamycin treatment is unpredictable and rapamycin is largely ineffective as a monotherapy (3). In particular, it has been shown that post-surgical, maintenance rapamycin treatment led to PI3K/Akt activation in glioblastoma patients, and this activation was associated with shorter time-to-progression (4). We asked whether PI3K hyperactivation induced by Grb10 knockdown (which phenotypically mimics rapamycin-induced Grb10 depletion) would contribute to the survival of cells undergoing stress-induced apoptosis. In response to either staurosporine or etoposide, reduced caspase 3 cleavage was observed in Grb10 knockdown cells compared to control cells. These results indicate that Grb10 depletion is sufficient to protect cells from apoptosis (FIGS. 4B and 10E). Combined with the recent finding that rapamycin can protect cells from energy stress-induced death (18), these results provide a plausible mechanism as to why the effect of rapamycin therapy is cytostatic rather than cytotoxic in some types of cancers and suggest a complete understanding of the feedback inhibition control will be critical in designing combination therapy involving rapamycin and DNA-damaging reagents or reagents targeting metabolic pathways.
To explore the role of Grb10 in cancer progression, we performed a comprehensive meta-analysis of GRB10 expression based on published microarray data. GRB10 expression is significantly down-regulated in many tumor types compared to their normal tissue counterparts (FIG. 4C). The most profound down-regulation was observed in infiltrating bladder urothelial carcinoma, glioblastoma, breast, myeloma, prostate and pancreatic cancers. Given that loss of Grb10 results in a dramatic activation of the PI3K/Akt pathway (FIG. 4A), we performed correlation analysis between variations in the expression of GRB10 and PTEN, a known negative regulator of the PI3K/Akt pathway and tumor suppressor gene. While both GRB10 and PTEN are ubiquitously down-regulated, in all cases there was a significantly (p<0.05) negative correlation between GRB10 and PTEN expression. The most striking differences were observed in breast carcinoma (Pearson correlation coefficient=−0.7, p=0.02) and myeloma (Pearson correlation coefficient=−0.82, p=0.001) (FIG. 4D). The data suggest that GRB10 loss provides an alternative mechanism of PI3K/Akt activation when PTEN expression is retained. It is particularly compelling that this correlation is only observed in tumor samples but not the normal tissue controls (FIG. 4E). It has been previously reported that PIK3CA mutations and PTEN loss are mutually exclusive in breast cancer (19), suggesting that an increased level of PIP₃resulting from of genetic alteration of either PIK3CA or PTEN relieves selective pressure targeting the other gene. Similarly, Grb10 loss, which results in PI3K hyperactivation, might provide the cells with growth and survival advantages that are redundant with respect to PTEN loss-of-function, suggesting that Grb10 might be a novel tumor suppressor that is regulated by mTORC1. These data point to the exciting therapeutic prospects of targeting Grb10 stability in cancer therapy.

Materials and Methods

Cells and Reagents

Human embryonic kidney (HEK) 293E cells, immortalized wild-type mouse embryonic fibroblast (MEF) cells and TSC2−/− MEFs (a kind gift from David Kwiatkowski, Brigham and Women's Hospital) were maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum. In collaboration with Millipore Inc., we generated anti-phospho-5501/S503-Grb10 antibodies. Anti-mTOR, anti-phospho-mTOR (S2481), anti-Grb10 (human), anti-phospho-Akt (S473), anti-phospho-Akt (T308), anti-Akt, anti-S6K, anti-phospho-S6K (T389), anti-IRS2, anti-PARP, anti-caspase 3, anti-4EBP, anti-4EBP (T37/T46), anti-phospho-ribosomal protein S6 (S235/S236), and anti-ribosomal protein S6 antibodies were obtained from Cell Signaling Technology. Anti-phospho-ERK1/2 antibody, insulin, Phorbol Myristate Acetate (PMA), Epidermal Growth Factor (EGF) and polybrene were purchased from Sigma. Anti-Grb10 (mouse), anti-phospho-IRS (Y612), anti-p85 and anti-p110 of PI3K antibodies were purchased from Santa Cruz, Invitrogen, Millipore and BD, respectively. ERK1/2 antibody and anti-HA antibody were prepared in the lab. LY294002 and AktVIII inhibitor were purchased from Calbiochem. Lipofectamine 2000 was purchased from Invitrogen. Torin was kindly provided by Nathanael Gray (Dana Farber Cancer Institute).

SILAC Cell Culture

TSC2−/− MEFs were used in the rapamycin screen due to constitutive hyperactivation of mTORC1 signaling in this cell line. Cells were grown in light ([¹²C₆ ¹⁴N₂]Lys, [¹²C₆ ¹⁴N₄]Arg) and heavy ([¹³C₆ ¹⁵N₂]Lys, [¹³C₆ ¹⁵N₄]Arg) DMEM (Cambridge Isotope Labs), respectively. Both light and heavy DMEM were supplemented with 10% dialyzed FBS (Invitrogen). Cells were serum-deprived for 17 hours and cells were cultured inheavy media were treated with 20 nM rapamycin for two hours. We performed two biological replicates of this experimental design with cross-labeling (swapping the labeled state of the rapamycin-treated cells). For the purposes of illustration, the data for biological replicate #2 is presented in FIGS. 1A and 1D.
The Ku-0063794 screen was performed using wild-type (WT) MEFs. Cells were grown in the aforementioned SILAC media. Both the light and heavy cells were starved of serum for 17 hrs. The light cells were treated with 20 nM rapamycin for 2 hrs, while the heavy cells were treated with a combination of 20 nM rapamycin and 2 μM Ku-0063794 for 2 hrs. Both the light and heavy cells were then stimulated with 100 nM insulin for 15 min.

Sample Preparation for Mass Spectrometric Analysis

The heavy and light cells were lysed in urea buffer (8 M urea, 20 mM HEPES pH 7.0, 75 mM β-glycerolphosphate, 1 mM sodium vanadate, 1 mM DTT and 1.5 mM EGTA) and the lysates were combined at a 1:1 ratio. Lysates were reduced by adding DTT to a final concentration of 3 mM, followed by incubation at room temperature for 20 min. Cysteines were alkylated by adding iodoacetamide to a final concentration of 50 mM, followed by incubation in the dark for 20 min. The lysates were diluted to a final concentration of 2 M urea by addition of 100 mM NH₄OAC and were digested overnight with sequencing-grade trypsin (Promega) at a 1:100 (enzyme:substrate) ratio. Digestion was quenched by addition of trifluoroacetic acid to a final concentration of 0.1% and precipitates were removed by centrifugation at 4,000 rpm for 30 min. Peptides were desalted on SepPak C18 columns (Waters) according to manufacturer's instructions.
Phosphopeptides were enriched by SCX-IMAC (27). Briefly, lyophilized peptides were resuspended in 500 μl SCX buffer A (5 mM KH₂PO₄, pH 2.65, 30% acetonitrile) and injected onto a SCX column (Polysulfoethyl aspartamide, 9.4 mm×200 mm, 5 μM particle size, 200 Å pore size, PolyLC). Gradient was developed over 35 min ranging from 0% to 21% buffer B (5 mM KH₂PO₄, pH 2.65, 30% acetonitrile, 350 mM KCl) at a flow rate of 2 ml/min. Twelve fractions were collected and lyophilized. Peptides were then desalted using SepPak C18 columns and were subjected to IMAC (Sigma) for phosphopeptide enrichment. The eluate was further desalted using STAGE tips (28) and lyophilized.

Mass Spectrometry Analysis and Data Processing

The rapamycin screen samples were analyzed by LC-MS/MS on an LTQ-Orbitrap mass spectrometer (Thermo, San Jose, Calif.) using the top ten method. The Ku-0063794 screen samples were analyzed on an LTQ-Velos mass spectrometer (Thermo Fischer Scientific, San Jose, Calif.) using the top twenty method. MS/MS spectra were searched against a composite database of the mouse IPI protein database (Version 3.60) and its reversed complement using the Sequest algorithm. Search parameters allowed for a static modification of 57.02146 Da for Cys and a dynamic modification of phosphorylation (79.96633 Da) on Ser, Thr and Tyr, oxidation (15.99491 Da) on Met, stable isotope (10.00827 Da) and (8.01420 Da) on Arg and Lys, respectively. Search results were filtered to include <1% matches to the reverse data base by the linear discriminator function (Huttlin et al., manuscript in preparation) using parameters including Xcorr, dCN, missed cleavage, charge state (exclude 1+ peptides), mass accuracy, peptide length and fraction of ions matched to MS/MS spectra. Phosphorylation site localization was assessed by the Ascore algorithm (29) based on the observation of phosphorylation-specific fragment ions and peptide quantification was performed by using the Vista algorithm (30, 31).
We further filter the peptides according to the following criteria for quantitation of the peptide abundance changes: (1) Vista confidence score must be at least 85, (2) signal to noise ratio (S/N)≧3 for both the heavy and light peptides, (3) in the cases where one of the isotopic species has an S/N of ≦3, S/N of the other was required to be ≧5, and (4) in the cases where only the heavy or light version of a peptide was found, we reported the peak S/N ratio, or its inverse, as a proxy for relative abundance measurement. For such peptides, we also required S/N of at ≧5 for the observed species.

Plasmids

The cDNA for human Grb10 (NCBI gene symbol GRB10; Gene ID: 2887) was obtained from Invitrogen and amplified by PCR. The product was subcloned into (1) the BamH I and EcoR I sites of pKH3, (2) the BamH I and EcoR I sites of pGEX-4T-3 or (3) the Hind III and EcoR I sites of pLPCX. The Grb10 point mutant constructs were generated using the QuickChange site-directed mutagenesis kit (Stratagene). pRK5-Myc-raptor andpRK5-Myc-Rictor were kindly provided by David Sabatini (MIT). Lentiviral plasmids (Δ8.9 and VSVG) were kind gifts from Andrew Kung (Dana Farber Cancer Institute) and David Baltimore (California Institute of Technology).

Immunoprecipitation

Cells were extracted with lysis buffer A (40 mM HEPES, pH 7.5, 120 mM NaCl, 1 mM EDTA, 10 mM β-glycerophosphate, 50 mM NaF, 2 mM phenylmethylsulfonyl fluoride, 2 mg/ml aprotinin, 2 mg/ml leupeptin, and 1 mg/ml pepstatin, 1 mM DTT) containing 1% Triton X-100, 1% NP-40, or 0.2% CHAPS. After centrifugation, supernatants were collected and pre-cleared for 1 h with protein A- and G-Sepharose beads (GE Healthcare Biosciences). After centrifugation at 3,000 rpm for 5 min, the supernatants were incubated with the antibody at 4° C. for 2 h, and then incubated with protein A- and G-Sepharose for an additional hour. Beads were washed four times with the lysis buffer and eluted in 2× reducing sample buffer.
Mammalian Lentiviral shRNAs
Lentiviral short hairpin RNA (shRNA) expression vectors were a kind gift from William Hahn (Dana Farber Cancer Institute). To generate the lentiviruses, shRNA plasmids were co-transfected into HEK293TD cells along with packaging (Δ8.9) and envelope (VSVG) expression plasmids using lipofectamine 2000 (Invitrogen). Two days after transfection, viral supernatants were harvested and filtered. Recipient cells were infected in the presence of a serum-containing medium supplemented with 8 μg/ml polybrene. Following infection for 36 h, cells were treated with 2.0 μg/ml puromycin (Sigma) and cell lines that stably expressed the shRNAs were selected. Knockdown efficiencies were examined by immunoblot assay using antibodies against the target protein.

Immunoblot Analysis

For immunoblot analysis, the cells were extracted in lysis buffer (20 mM HEPES (pH 7.5), 1% Triton X-100, 150 mM NaCl, 10 mM EDTA, 1 mM EGTA, 1 mM sodium orthovanadate, 1 mM NaF, 2 mM phenylmethylsulfonyl fluoride, 2 mg/ml aprotinin, 2 mg/ml leupeptin, and 1 mg/ml pepstatin), and extracts were mixed with the 5× reducing buffer (60 mM Tris-HCl, pH 6.8, 25% glycerol, 2% SDS, 14.4 mM 2-mercaptoethanol, 0.1% bromophenol blue). Samples were boiled for 5 min and subject to electrophoresis using the standard SDS-PAGE method. Proteins were then transferred to a nitrocellulose membrane (Whatman). The membranes were blocked with a TBST buffer (25 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.05% Tween 20) containing 3% nonfat dried milk, and probed overnight with primary antibodies at 4° C. and for 1 h at RT with peroxidase-conjugated secondary antibodies. Blots were developed using enhanced chemiluminescence, exposed on autoradiograph film and developed using standard methods.

Recombinant Protein Purification

For the purification of GST-tagged proteins, plasmids were transformed into Escherichia coli strain BL21 (DE3), and purified to homogeneity from crude lysates using glutathione-sepharose beads (GE Healthcare) according to the manufacture's protocol. Briefly, protein production was initiated by adding isopropyl-D-thiogalactopyranoside (Sigma) to the cultures. Bacteria were collected by centrifugation, resuspended in PBS and lysed by sonication. After centrifugation at 13,000 rpm for 15 min, the supernatant was incubated with glutathione-sepharose beads for 1 h. The beads were washed with PBS three times and the recombinant protein was eluted with PBS containing 20 mM reduced glutathione. Proteins were dialyzed against PBS and stored at −80° C. until use.

Quantitative RT-PCR Analysis

Total cellular RNA was purified from cultured cells using the RNeasy mini kit (Qiagen) following the manufacturer's protocol. For quantitative real-time PCR (qRT-PCR), RNA was reverse-transcribed using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems) according to the manufacturer's instructions. The resulting cDNA was analyzed by qRT-PCR using the QuantiTect SYBR Green qPCR System (Qiagen). A QuantiTect Primer Assay for mouse Grb10 was used to amplify the target gene, while the β-acting primers (β-actin forward, ACCCAGATCATGTTTGAGACCT (SEQ ID NO: 25); and β-actin reverse, GCAGTAATCTCCTTCTGCATCC (SEQ ID NO: 26)) were used as a normalization control. All reactions were run on an ABI 7900HT Fast Real-Time PCR instrument with a 15 min hot start at 95° C. followed by 40 cycles of a 3-step thermocycling program: denaturation: 15 s at 94° C., annealing: 30 s at 55° C. and extension: 30 s at 70° C. Melting curve analysis was performed at the end of every run to ensure that a single PCR product of the expected melting temperature was produced in a given well. A total of 3 biological replicates×4 technical replicates/biological replicate were performed for each treatment group. Data analysis utilized the comparative C_tmethod (ΔΔC_tmethod).

Analysis of GRB10 and PTEN Expression in Human Samples.

Microarray expression data from six independent data sets corresponding to patient samples from bladder (GSE3167), glioblastoma (GSE4536), breast (GSE5764), myeloma (GSE5900), pancreatic (GSE1542) carcinoma and matching normal tissues were downloaded from Gene Expression Omnibus (www.nebi.nlm.nih.gov/geo/) and for prostate carcinoma from the Broad Institute cancer program datasets (www.broadinstitute.org/cgi-bin/cancer/datasets.cgi). The correlation in the gene expression between GRB10 and PTEN was performed using Pearson's correlation coefficient analysis and the samples were clustered using the Euclidean distance metric and Ward's linkage algorithm.

REFERENCES

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INCORPORATION BY REFERENCE

All publications, patents and sequence database entries mentioned herein, including those items listed below and in the databases and tables provided or referred to herein, are hereby incorporated by reference in their entirety for disclosure of the relevant subject matter indicated, as if each individual database entry, publication, or patent was specifically and individually indicated to be incorporated by reference. The specification provides a number of International Protein Index (IPI) accession numbers, starting with IPI followed by a number. International Protein Index database entries identified by IPI accession number in the specification are incorporated by reference for disclosure of the respective protein sequence and accompanying protein information. The IPI database can be accessed at the European Bioinformatics Institute homepage (www.ebi.ac.uk/), for example, at (www.ebi.ac.uk/IPI/IPIhelp.html). In case of conflict, the present application, including any definitions herein, will control.

EQUIVALENTS AND SCOPE

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. The scope of the present invention is not intended to be limited to the above description, but rather is as set forth in the appended claims.
In the claims articles such as “a,” “an,” and “the” may mean one or more than one unless indicated to the contrary or otherwise evident from the context. Claims or descriptions that include “or” between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process unless indicated to the contrary or otherwise evident from the context. The invention includes embodiments in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process. The invention also includes embodiments in which more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process. Furthermore, it is to be understood that the invention encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, descriptive terms, etc., from one or more of the claims or from relevant portions of the description is introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Furthermore, where the claims recite a composition, it is to be understood that methods of using the composition for any of the purposes disclosed herein are included, and methods of making the composition according to any of the methods of making disclosed herein or other methods known in the art are included, unless otherwise indicated or unless it would be evident to one of ordinary skill in the art that a contradiction or inconsistency would arise. For example, it is to be understood that any of the compositions of the invention can be used for vocal cord repair or other soft tissue repair or augmentation. It is also to be understood that any of the compositions made according to the methods for preparing compositions disclosed herein can be used for vocal cord repair or other soft tissue repair or augmentation. In addition, the invention encompasses compositions made according to any of the methods for preparing compositions disclosed herein.
Where elements are presented as lists, e.g., in Markush group format, it is to be understood that each subgroup of the elements is also disclosed, and any element(s) can be removed from the group. It is also noted that the term “comprising” is intended to be open and permits the inclusion of additional elements or steps. It should be understood that, in general, where the invention, or aspects of the invention, is/are referred to as comprising particular elements, features, steps, etc., certain embodiments of the invention or aspects of the invention consist, or consist essentially of, such elements, features, steps, etc. For purposes of simplicity those embodiments have not been specifically set forth in haec verba herein. Thus for each embodiment of the invention that comprises one or more elements, features, steps, etc., the invention also provides embodiments that consist or consist essentially of those elements, features, steps, etc.
Where ranges are given, endpoints are included. Furthermore, it is to be understood that unless otherwise indicated or otherwise evident from the context and/or the understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value within the stated ranges in different embodiments of the invention, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise. It is also to be understood that unless otherwise indicated or otherwise evident from the context and/or the understanding of one of ordinary skill in the art, values expressed as ranges can assume any subrange within the given range, wherein the endpoints of the subrange are expressed to the same degree of accuracy as the tenth of the unit of the lower limit of the range.
In addition, it is to be understood that any particular embodiment of the present invention may be explicitly excluded from any one or more of the claims. Any embodiment, element, feature, application, or aspect of the compositions and/or methods of the invention, can be excluded from any one or more claims. For purposes of brevity, all of the embodiments in which one or more element, feature, purpose, or aspect is excluded are not set forth explicitly herein.

TABLES

The Tables provided below are referred to in the specification and the claims. Because some of the tables are lengthy, they are provided in this subsection.

TABLE 1

Gene Symbol	Annotation

1110013L07Rik	Putative uncharacterized protein
1600027N09Rik	RIKEN cDNA 1600027N09 gene
2610110G12Rik	Isoform 1 of UPF0635 protein C6orf134 homolog
Aak1	Isoform 2 of AP2-associated protein kinase 1
Adamts2	A disintegrin and metalloproteinase with thrombospondin motifs 2
Ahctf1	AT-hook-containing transcription factor 1
Ahnak	AHNAK nucleoprotein isoform 1
Aim1l	Absent in melanoma 1-like
Akt1s1	Proline-rich AKT1 substrate 1
Alkbh6	24 kDa protein
Arhgap17	Isoform 1 of Rho GTPase-activating protein 17
Atg2a	Autophagy-related protein 2 homolog A
B230208H17Rik	Putative GTP-binding protein Parf
Bat2l	Isoform 1 of Protein BAT2-like
Bcas3	Isoform 1 of Breast carcinoma-amplified sequence 3 homolog
Bclaf1	Isoform 2 of Bcl-2-associated transcription factor 1
Bcr	Breakpoint cluster region protein
Bmp2k	Isoform 1 of BMP-2-inducible protein kinase
Bod1l	biorientation of chromosomes in cell division 1-like
C130092O11Rik	Isoform 1 of Uncharacterized protein KIAA1680
Cabin1	calcineurin binding protein 1
Carhspl	Calcium-regulated heat stable protein 1
Ccdc6	coiled-coil domain containing 6
Ccnl1	Isoform 1 of Cyclin-L1
Cd2ap	CD2-associated protein
Cdgap	Cdc42 GTPase-activating protein
Chd1	Chromodomain-helicase-DNA-binding protein 1
Crkrs	Isoform 2 of Cell division cycle 2-related protein kinase 7
Cttn	Src substrate cortactin
Cux1	cut-like homeobox 1 isoform a
D6Wsu116e	Isoform 1 of Protein FAM21
D830015G02Rik	Putative uncharacterized protein
Dab2	Isoform p93 of Disabled homolog 2
Ddx21	Nucleolar RNA helicase 2
Dennd4a	hypothetical protein LOC102442
Dhx15	Putative pre-mRNA-splicing factor ATP-dependent RNA helicase DHX15
Dnajc2	DnaJ homolog subfamily C member 2
Dock11	Dedicator of cytokinesis protein 11
Edc3	Enhancer of mRNA-decapping protein 3
Edc4	Isoform 1 of Enhancer of mRNA-decapping protein 4
Eef2k	Elongation factor 2 kinase
Ehbp1	Isoform 2 of EH domain-binding protein 1
Ehmt2	Isoform 1 of Histone-lysine N-methyltransferase H3 lysine-9 specific 3
Eif3a	Eukaryotic translation initiation factor 3 subunit A
Eif4b	Eukaryotic translation initiation factor 4B
Eif4ebp1	Eukaryotic translation initiation factor 4E-binding protein 1
Eif4ebp2	Eukaryotic translation initiation factor 4E-binding protein 2
Eif4g1	Isoform 1 of Eukaryotic translation initiation factor 4 gamma 1
Eif5; LOC100047658	Eukaryotic translation initiation factor 5
Emg1	Probable ribosome biogenesis protein NEP1
Eps8l2	Isoform 1 of Epidermal growth factor receptor kinase substrate 8-like protein 2
Epyc	Epiphycan
Erc1; LOC100048600	Isoform 1 of ELKS/RAB6-interacting/CAST family member 1
Ext1	Exostosin-1
Fbxw9	F-box and WD-40 domain protein 9
Fkbp15	Isoform B of FK506-binding protein 15
FLNA	Filamin-A
Fmnl3	Isoform 1 of Formin-like protein 3
Foxk1	Forkhead box protein K1
Foxk2	Isoform 1 of Forkhead box protein K2
Fxr1	Isoform E of Fragile X mental retardation syndrome-related protein 1
Gbf1	Golgi-specific brefeldin A-resistance factor 1
Gm13099	Novel protein similar to preferentially expressed antigen in melanoma-like family
Gm13697	Novel protein containing MIF4G and MA3 domains
Gm14085	Novel protein similar to solute carrier family 28 (Sodium-coupled nucleoside
	transporter) member 2
Gm6988	similar to hCG1640785
Gm9757	Putative uncharacterized protein
Gon4l	RIKEN cDNA 5830417110 gene
Grb10	Isoform 3 of Growth factor receptor-bound protein 10
Gsk3b	Glycogen synthase kinase-3 beta
Gtf2f1	General transcription factor IIF subunit 1
Hdgfrp2	Isoform 3 of Hepatoma-derived growth factor-related protein 2
Herc1	hect (homologous to the E6-AP (UBE3A) carboxyl terminus) domain and RCC1
	(CHC1)-like domain (RLD) 1
Hisppd1	140 kDa protein
Hn1l	Hematological and neurological expressed 1-like protein
Ibtk	Isoform 2 of Inhibitor of Bruton tyrosine kinase
Inf2	Isoform 1 of Inverted formin-2
Iqsec1	IQ motif and Sec7 domain 1 isoform b
Irs2	Insulin receptor substrate 2
Iws1	Isoform 1 of Protein IWS1 homolog
Junb	Transcription factor jun-B
Kdm3b	Isoform 2 of Lysine-specific demethylase 3B
Kdm6a	Isoform 1 of Lysine-specific demethylase 6A
Ktn1	Isoform 1 of Kinectin
Larp1	Isoform 1 of La-related protein 1
Larp4	Putative uncharacterized protein
Larp7	Isoform 1 of La-related protein 7
Ldhd	22 kDa protein
Lin9	lsoform 2 of Lin-9 homolog
Llgl1	lethal giant larvae homolog 1 isoform 1
LOC100048123; Akt2	RAC-beta serine/threonine-protein kinase
LOC100048559; Sfrs1	Isoform 1 of Splicing factor arginine/serine-rich 1
Luc7l2	Isoform 1 of Putative RNA-binding protein Luc7-like 2
Macf1	Isoform 3 of Microtubule-actin cross-linking factor 1
Med1	Isoform 4 of Mediator of RNA polymerase II transcription subunit 1
Megf11	Isoform 4 of Multiple epidermal growth factor-like domains 11
Mett10d	Isoform 1 of Putative methyltransferase METT10D
Mib1	E3 ubiquitin-protein ligase MIB1
Micall1	Isoform 1 of MICAL-like protein 1
Mll2	similar to myeloid/lymphoid or mixed-lineage leukemia 2
Mllt4	Isoform 3 of Afadin
Mogat1	2-acylglycerol O-acyltransferase 1
Mtap1b	Microtubule-associated protein 1B
Mtor	Isoform 1 of FKBP12-rapamycin complex-associated protein
Myef2	Isoform 2 of Myelin expression factor 2
Myo5a	Myosin-Va
Nacc1	Nucleus accumbens-associated protein 1
Ndrg3	Protein NDRG3
Nfic	Isoform 1 of Nuclear factor 1 C-type
Npm1	Nucleophosmin
Numa1	Nuclear mitotic apparatus protein 1
Palm	Isoform 1 of Paralemmin
Patl1	Protein PAT1 homolog 1
Pbx2	Pre-B-cell leukemia transcription factor 2
Pcbp1	Poly(rC)-binding protein 1
Pcbp2	Isoform 1 of Poly(rC)-binding protein 2
Pcm1	Isoform 1 of Pericentriolar material 1 protein
Pdcd11	Protein RRP5 homolog
Pds5b	Isoform 1 of Sister chromatid cohesion protein PDS5 homolog B
Peg3	Isoform 1 of Paternally-expressed gene 3 protein
Pgrmc2	Membrane-associated progesterone receptor component 2
Phf3	PHD finger protein 3
Phldb1	Isoform 2 of Pleckstrin homology-like domain family B member 1
Phldb2	lsoform 1 of Pleckstrin homology-like domain family B member 2
Pi4k2a	Phosphatidylinositol 4-kinase type 2-alpha
Pkn2	Isoform 1 of Serine/threonine-protein kinase N2
Pla2g4a	Cytosolic phospholipase A2
Plekhm1	Pleckstrin homology domain-containing family M member 1
Pom121	Nuclear envelope pore membrane protein POM 121
Ppfibp2	Isoform 4 of Liprin-beta-2
Ppp1r12a	MCG122391 isoform CRA_e
Prkd2	Serine/threonine-protein kinase D2
Ptk2	Isoform 1 of Focal adhesion kinase 1
Qsox2	Isoform 3 of Sulfhydryl oxidase 2
Rab1	Ras-related protein Rab-1A
Ranbp10	Ran-binding protein 10
Ranbp9	RAN binding protein 9
Rb1	Retinoblastoma-associated protein
Rfc1	Rfc1 protein
Rictor	Isoform 1 of rapamycin-insensitive companion of mTOR
Rnf19b	IBR domain containing 3
Rps6	29 kDa protein
Rps6kb1	Isoform Alpha I of Ribosomal protein S6 kinase beta-1
RsI1d1	Putative uncharacterized protein
rsp6	29 kDa protein
Sap30	Histone deacetylase complex subunit SAP30
Sbno1	Isoform 2 of Protein strawberry notch homolog 1
Scrib	Isoform 1 of Protein LAP4
Serbp1	Isoform 1 of Plasminogen activator inhibitor 1 RNA-binding protein
Setd1a	SET domain containing 1A
Sfrs18	splicing factor arginine/serine-rich 18
Sfrs8	splicing factor arginine/serine-rich 8
Sgta	Isoform 1 of Small glutamine-rich tetratricopeptide repeat-containing protein alpha
Slc4a1ap	solute carrier family 4 (anion exchanger) member 1 adaptor protein
Smarca4	Putative uncharacterized protein
Smarcad1	Isoform 1 of SWI/SNF-related matrix-associated actin-dependent regulator of
	chromatin subfamily A containing DEAD/H box 1
Smarcc2	Isoform 2 of SWI/SNF complex subunit SMARCC2
Snx17	Sorting nexin-17
Snx30	Sorting nexin-30
Sorbs3	Vinexin
Spna2	Isoform 2 of Spectrin alpha chain brain
Sqstm1	Isoform 1 of Sequestosome-1
Srpk2	serine/arginine-rich protein-specific kinase 2
Srrm1	Isoform 2 of Serine/arginine repetitive matrix protein 1
Srrm2	Isoform 3 of Serine/arginine repetitive matrix protein 2
St5	Isoform 1 of Suppression of tumorigenicity 5
Strn3	Striatin-3
Syap1	Synapse-associated protein 1
Tbc1d4	140 kDa protein
Tcfeb	Transcription factor EB
Tinf2	Putative uncharacterized protein
Tjp2	Tight junction protein ZO-2
Tmem106b	Transmembrane protein 106B isoform CRA_b
Tmpo	Isoform Beta of Lamina-associated polypeptide 2 isoforms beta/delta/epsilon/gamma
Tnc	Isoform 1 of Tenascin
Tns1	tensin 1
Tox4	TOX high mobility group box family member 4
Trim28	Isoform 1 of Transcription intermediary factor 1-beta
Trip10	Isoform 3 of Cdc42-interacting protein 4
Ubxn7	UBX domain-containing protein 7
Ulk1	Putative uncharacterized protein
Usp24	Isoform 1 of Ubiquitin carboxyl-terminal hydrolase 24
Usp36	Ubiquitin specific peptidase 36
Uvrag	UV radiation resistance associated
Vwa5b1	von Willebrand factor A domain-containing protein 5B1
Wdr91	WD repeat-containing protein 91
Wnk1	Serine/threonine-protein kinase WNK1
Zc3h4	Isoform 2 of Zinc finger CCCH domain-containing protein 4
Zc3hc1	Isoform 1 of Nuclear-interacting partner of ALK
Zeb2	Zinc finger E-box-binding homeobox 2
Zfp106	Isoform 1 of Zinc finger protein 106
Zfp516	Zinc finger protein 516
Znrf2	E3 ubiquitin-protein ligase ZNRF2

Table 1. Names and annotations of proteins identified to harbor phosphopeptides whose intensities decrease after rapamycin treatment (rapamycin screen).
Note
that the rapamycin screen included two biological replicates. For the first replicate, the light cells were controls whereas the heavy cells were treated with rapamycin. For the second replicate experiment, the light cells were treated with rapamycin, and the heavy cells were controls. Table 1 includes all unique proteins identified in both replicates. Protein sequences and phosphorylation sites can be identified from the identifiers provided in the tables and database provided herein.

TABLE 2

Gene Symbol	Annotation

Myo18a	Isoform 4 of Myosin-XVIIIa
Brd2	Isoform 2 of Bromodomain-containing protein 2
6330577E15Rik	Uncharacterized protein C10orf78 homolog
Best3	Bestrophin-3
Larp1	Isoform 1 of La-related protein 1
Fam62c	Isoform 1 of Extended synaptotagmin-3
Mdn1	Midasin homolog
Rtn4	Isoform 2 of Reticulon-4
Pcdh24	Pcdh24 protein
Synj1	similar to mKIAA0910 protein
Zfp318	zinc finger protein 318 isoform 1
Akt1s1	Proline-rich AKT1 substrate 1
Alpk3	myocyte induction differentiation originator
Zc3h14	Isoform 2 of Zinc finger CCCH domain-containing
	protein 14
Edc4	Isoform 1 of Enhancer of mRNA-decapping
	protein 4
Srrm2	Isoform 3 of Serine/arginine repetitive matrix
	protein 2
Kif20b	Isoform 1 of M-phase phosphoprotein 1
1110007A13Rik	UPF0557 protein C10orf119 homolog
Pi4k2a	Phosphatidylinositol 4-kinase type 2-alpha
Top2b	DNA topoisomerase 2-beta
BC021381	Isoform 2 of Uncharacterized protein KIAA1931
Bbx	Isoform 1 of HMG box transcription factor BBX
Vcl	Vinculin
Eif4ebp1	Eukaryotic translation initiation factor 4E-binding
	protein 1
Rrp15	RRP15-like protein
D10Wsu102e	Uncharacterized protein C12orf45 homolog
Braf	Isoform 1 of B-Raf proto-oncogene serine/
	threonine-protein kinase
Atg2b	Isoform 1 of Autophagy-related protein 2
	homolog B
Npm1	Nucleophosmin
Pcsk5	proprotein convertase subtilisin/kexin type 5
Mtap1b	Microtubule-associated protein 1B
Ercc6l	DNA excision repair protein ERCC-6-like
Serinc1	Serine incorporator 1
Klf3; LOC100046855	Krueppel-like factor 3
Smarca4	Putative uncharacterized protein
Aak1	Uncharacterized protein FLJ45252 homolog
Eif4ebp2	Eukaryotic translation initiation factor 4E-binding
	protein 2
Ndrg1	Protein NDRG1
Melk	Maternal embryonic leucine zipper kinase
Arhgef17	Isoform 1 of Rho guanine nucleotide exchange
	factor 17
Grit	Isoform 2 of Rho/Cdc42/Rac GTPase-activating
	protein RICS
Mdc1	mediator of DNA damage checkpoint 1
Nfkb2	NF-kB2 splice variant 4
Pcm1	Isoform 1 of Pericentriolar material 1 protein
Mybbp1a	Myb-binding protein 1A
Sf3b1	Splicing factor 3B subunit 1
Atrx	Transcriptional regulator ATRX
Ccdc88a	Isoform 2 of Girdin
Baz1b	Isoform 1 of Tyrosine-protein kinase BAZ1B
Nedd4l	Isoform 3 of E3 ubiquitin-protein ligase NEDD4-
	like
Orc6l	Origin recognition complex subunit 6
Trp53bp1	Transformation related protein 53 binding protein 1
Map3k2	Mitogen-activated protein kinase kinase kinase 2
Hectd2	Hectd2 protein
Usp10	Ubiquitin carboxyl-terminal hydrolase 10
D830031N03Rik	similar to mKIAA0754 protein
Nck1	non-catalytic region of tyrosine kinase adaptor
	protein 1
Exosc9	Exosome complex exonuclease RRP45
Dap	Death-associated protein 1
Rps6kb1	Isoform Alpha I of Ribosomal protein S6 kinase
	beta-1
Lmna	Isoform C2 of Lamin-A/C
Sltm	Isoform 1 of SAFB-like transcription modulator
Sh3pxd2a	Isoform 1 of SH3 and PX domain-containing
	protein 2A
Flnc	Isoform 1 of Filamin-C
Oxr1	Isoform 2 of Oxidation resistance protein 1
Rin2	Isoform 1 of Ras and Rab interactor 2
Nek9	Serine/threonine-protein kinase Nek9
Pebp1	Phosphatidylethanolamine-binding protein 1
Pop1	Processing of 1 ribonuclease P/MRP family
Serhl	Serine hydrolase-like protein
Epb4.1l3	Isoform 1 of Band 4.1-like protein 3
Hnrpll	Isoform 1 of Heterogeneous nuclear
	ribonucleoprotein L-like
Samhd1	SAM domain and HD domain-containing protein 1
Zfp828	Zinc finger protein 828
Larp7	Isoform 1 of La-related protein 7
Myc	myc proto-oncogene protein
Myo9a	Isoform 2 of Myosin-IXa
Gsk3b	Glycogen synthase kinase-3 beta
Zfp395	zinc finger protein 395
Bend3	BEN domain-containing protein 3
Akap12	Isoform 1 of A-kinase anchor protein 12
Eif4g1	Isoform 1 of Eukaryotic translation initiation factor
	4 gamma 1
Eif4b	Eukaryotic translation initiation factor 4B
Dock7	Isoform 2 of Dedicator of cytokinesis protein 7
Patl1	Protein PAT1 homolog 1
Slc7a11	Cystine/glutamate transporter
Myo9b	Isoform 1 of Myosin-IXb
Setd2	SET domain containing 2
Gphn	Gephyrin
Erf	ETS domain-containing transcription factor ERF
Spnb2	Isoform 2 of Spectrin beta chain brain 1
Phip	PH-interacting protein
Sdpr	Serum deprivation-response protein
Tcof1	Treacle protein
Pwp1	Periodic tryptophan protein 1 homolog
Rbl1	Isoform Long of Retinoblastoma-like protein 1
Eef1b2	Elongation factor 1-beta
Phactr4	Isoform 1 of Phosphatase and actin regulator 4
C230081A13Rik	Tyrosine-protein kinase-protein kinase SgK269
Ahnak2	Putative uncharacterized protein

Table 2. Names and annotations of proteins identified to harbor phosphopeptides whose intensities decrease after Ku-0063794 treatment (Ku-0063794 screen).
Note
that the light cells were treated with rapamycin and serve as controls whereas the heavy cells were treated with a combination of rapamycin and Ku-0063794. Protein sequences and phosphorylation sites can be identified from the identifiers provided in the tables and the database provided herein.

TABLE 3

		SEQ
		ID	Gene
IPI Reference	Peptide	NO	Name	Class

IPI00121418.1	K.DGEGPDNLEPACPLSLPLQGNHTAADMYLSPLRSPK.K	30	Rb1	1

IPI00221581.1	R.TGSESSQTGASATSGR.N	31	Eif4b	1

IPI00929786.1	R.TASISSSPSEGTPAVGSYGCTPQSLPK@.F	32	Larp1	1

IPI00929786.1	R.TASISSSPSEGTPAVGSYGCTPQSLPK@.F	33	Larp1	1

IPI00929786.1	R.TASISSSPSEGTPAVGS*YGCTPQSLPK@.F	34	Larp1	1

IPI00317401.6	R.AESPETSAVESTQSTPQK@.G	35	Pds5b	1

IPI00107958.1	K.TSDIFGS*PVTATAPLAHPNK@PK@.D	36	Hn1l	1

IPI00108454.2	R.R{circumflex over ( )}LSSLR{circumflex over ( )}.A	37	Rps6	1

IPI00225062.2	R.R{circumflex over ( )}SSSELSPEVVEK@.V	38	Srrm2	1

IPI00761759.1	R.SSSGSEHST*EGSVSLGDGPLSR{circumflex over ( )}.S	39	Larp4	1

IPI00761759.1	R.SSSGSEHST*EGSVSLGDGPLSR{circumflex over ( )}.S	40	Larp4	1

IPI00225062.2	R.RSSSELSPEVVEK.V	41	Srrm2	1

IPI00122594.4	R.TTPLASPSLSPGR{circumflex over ( )}.S	42	Ahctf1	1

IPI00136107.1	R.THSTSSS*IGSGESPFSR{circumflex over ( )}.S	43	Ndrg3	1

IPI00136107.1	R.THSTSSSIGSGESPFSR{circumflex over ( )}.S	44	Ndrg3	1

IPI00136107.1	R.THSTSSSIGSGESPFSR{circumflex over ( )}.S	45	Ndrg3	1

IPI00136107.1	R.THS*TSSSIGSGESPFSR{circumflex over ( )}.S	46	Ndrg3	1

IPI00136107.1	R.THSTS*SSIGSGESPFSR{circumflex over ( )}.S	47	Ndrg3	1

IPI00136107.1	R.THSTSSSIGSGESPFSR{circumflex over ( )}.S	48	Ndrg3	1

IPI00136107.1	R.THSTSSSIGSGESPFSR{circumflex over ( )}.S	49	Ndrg3	1

IPI00136107.1	R.THSTSSSIGSGESPFSR{circumflex over ( )}.S	50	Ndrg3	1

IPI00136107.1	R.THS*TSSSIGSGESPFSR{circumflex over ( )}.S	51	Ndrg3	1

IPI00551454.3	R.ER{circumflex over ( )}QES*ESEQELVNK@.R	52	Pdcd11	1

IPI00117229.3	R.TS*PAGGTWSSVVSGVPR.L	53	Atxn2	1

IPI00348442.1	R.SK@FDS*DEEDEDAENLEAVSSGK@.V	54	Sfrs18	1

IPI00556837.1	K.DTVIIVSEPSEDEESHDLPSVTR.R	55	Smarcad1	1

IPI00153986.2	K.GTSRPGTPSAEAASTSSTLR.A	56	Gtf2f1	1

IPI00225062.2	R.GCSPPKSPEKPPQSTSSESCPPS*PQPTK.V	57	Srrm2	1

IPI00128904.1	R.VMTIPYQPMPASS*PVICAGGQDR.C	58	Pcbp1	1

IPI00121251.7	R.LSTTPSPTNSLHEDGVDDFRR.Q	59	Tox4	1

IPI00320594.5	K.SQPHSSTSNQETSDSEMEMEAEHYPNGVLESVSTR{circumflex over ( )}.I	60	Ranbp10	1

IPI00808277.2	R.SAPASPNHAGVLSAHSSGAQTPESLS*R{circumflex over ( )}.E	61	Foxk2	1

IPI00137166.1	R.SR{circumflex over ( )}DATPPVSPINMEDQER{circumflex over ( )}.I	62	Junb	1

IPI00454104.1	K.TTEAPCSPGSQQPPSPDELPANVK@.Q	63	Scrib	1

IPI00225062.2	R.GCSPPK@SPEK@PPQSTSSESCPPS*PQPTK@.V	64	Srrm2	1

IPI00153986.2	K.GTSR{circumflex over ( )}PGTPSAEAASTSSTLR{circumflex over ( )}.A	65	Gtf2f1	1

IPI00753321.2	K.SVSETSEDK@K@DEESDEEEEEEEEEEPLGATTR{circumflex over ( )}.S	66	Bod1l	1

IPI00153986.2	K.GTSR{circumflex over ( )}PGTPSAEAASTSSTLR{circumflex over ( )}.A	67	Gtf2f1	1

IPI00153986.2	K.GTSR{circumflex over ( )}PGTPSAEAASTSSTLR{circumflex over ( )}.A	68	Gtf2f1	1

IPI00929786.1	R.TASISSSPSEGTPAVGSYGCT*PQSLPK@.F	69	Larp1	1

IPI00929786.1	R.TASISSSPSEGTPAVGSYGCTPQSLPK@.F	70	Larp1	1

IPI00130920.1	R.RSES*PFECK.N	71	Mtap1b	1

IPI00336973.2	K.GLNLDGTPALSTLGGFSPASK@PSSPR{circumflex over ( )}.E	72	Ccnl1	1

IPI00336973.2	K.GLNLDGTPALSTLGGFSPASKPSSPR.E	73	Ccnl1	1

IPI00336973.2	K.GLNLDGTPALSTLGGFSPASK@PS*SPR{circumflex over ( )}.E	74	Ccnl1	1

IPI00336973.2	K.GLNLDGTPALSTLGGFSPASK@PSSPR{circumflex over ( )}.E	75	Ccnl1	1

IPI00379844.4	R.TASEGDGGAAGGAGTAGGR{circumflex over ( )}PMSVAGSPLS*PGPVR{circumflex over ( )}.A	76	Irs2	1

IPI00336973.2	K.GLNLDGTPALSTLGGFSPASKPSSPR.E	77	Ccnl1	1

IPI00336973.2	K.GLNLDGTPALSTLGGFSPASKPSS*PR.E	78	Ccnl1	1

IPI00656285.2	R.SLVSPIPSPTGTISVPNSCPAS*PR{circumflex over ( )}.G	79	Foxk1	1

IPI00656285.2	R.SLVSPIPSPTGTISVPNSCPAS*PR.G	80	Foxk1	1

IPI00454104.1	K.TTEAPCSPGSQQPPSPDELPANVK@.Q	81	Scrib	1

IPI00753321.2	K.SVSETSEDK@K@DEESDEEEEEEEEEEPLGATTR{circumflex over ( )}.S	82	Bod1l	1

IPI00753321.2	K.SVSETSEDK@K@DEESDEEEEEEEEEEPLGATTR{circumflex over ( )}.S	83	Bod1l	1

IPI00313307.3	R.SYQNSPSSEDGIR{circumflex over ( )}PLPEYSTEK@.H	84	Med1	1

IPI00225062.2	R.GCSPPK@SPEK@PPQSTSS*ESCPPSPQPTK@.V	85	Srrm2	1

IPI00318938.6	R.NSPVAK@TPPK@DLPAIPGVTSPTSDEPPMQASQSQL	86	Eif4ebp1	1
	PSSPEDK@.R

IPI00130920.1	R.SLMS*SPEDLTK@DFEELK@AEEIDVAK@.D	87	Mtap1b	1

IPI00553798.2	R.SSEVVLSGDDEDYQR{circumflex over ( )}.I	88	Ahnak	1

IPI00129264.1	R.ATSR{circumflex over ( )}PINLGPSS*PNTEIHWTPYR{circumflex over ( )}.A	89	Sorbs3	1

IPI00129264.1	R.ATSRPINLGPS*SPNTEIHWTPYR.A	90	Sorbs3	1

IPI00123410.5	R.TIS*AQDTLAYATALLNEK@.E	91	Usp24	1

IPI00226441.2	R.LHYTPPLQSPITDGDPLLGQSPWR{circumflex over ( )}.S	92	Lin9	1

IPI00309059.7	R.STSPIIGSPPVR{circumflex over ( )}.A	93	Patl1	1

IPI00336713.1	R.CSPVPGLSSSPSGSPLHGK@.L	94	Bcas3	1

IPI00320905.7	R.LGEQGPEPGPTPPQTPTPPS*TPPLAK.Q	95	Arhgap17	1

IPI00309059.7	R.R{circumflex over ( )}STSPIIGS*PPVR{circumflex over ( )}.A	96	Patl1	1

IPI00309059.7	R.RSTSPIIGSPPVR.A	97	Patl1	1

IPI00309059.7	R.STSPIIGS*PPVR{circumflex over ( )}.A	98	Patl1	1

IPI00317599.3	K.SQEDEEEISTS*PGVSEFVSDAFDTCSLNQEDLRK.E	99	Syap1	1

IPI00317599.3	K.SQEDEEEIST*SPGVSEFVSDAFDTCSLNQEDLRK.E	100	Syap1	1

IPI00656285.2	R.SLVSPIPSPTGTISVPNS*CPASPR{circumflex over ( )}.G	101	Foxk1	1

IPI00676574.2	R.RVS*TDLPEGQDVYTAACNSVIHR.C	102	Herc1	1

IPI00458958.2	K.DWDK@ES*EGEEPAGGR{circumflex over ( )}.A	103	Rrp15	2

IPI00230719.8	R.R{circumflex over ( )}DSS*DDWEIPDGQITVGQR{circumflex over ( )}.I	104	Braf	2

IPI00318938.6	R.VALGDGVQLPPGDYSTTPGGTLFSTTPGGTR.I	105	Eif4ebp1	2

IPI00874995.2	R.AS*DDLGEPDVFATAPFR.S	106	Aak1	2

IPI00318938.6	R.VALGDGVQLPPGDYSTTPGGTLFSTTPGGTR{circumflex over ( )}.I	107	Eif4ebp1	2

IPI00318938.6	R.VALGDGVQLPPGDYSTTPGGTLFSTTPGGTR.I	108	Eif4ebp1	2

IPI00318938.6	R.VALGDGVQLPPGDYSTTPGGTLFSTTPGGTR{circumflex over ( )}.I	109	Eif4ebp1	2

IPI00323045.3	R.S*MDVDLNQAHMEDTPK@.K	110	Melk	2

IPI00623284.4	K.IWDPTPSHTPAGAATPGRGDTPGHATPGHGGATSSA	111	Sf3b1	2
	R.K

IPI00125960.1	R.TASGSSVTSLEGTR.S	112	Ndrg1	2

IPI00318938.6	R.R{circumflex over ( )}VALGDGVQLPPGDYSTTPGGTLFSTTPGGTR{circumflex over ( )}.I	113	Eif4ebp1	2

IPI00229571.1	R.ISSK@SPGHMVILNQTK@.G	114	Sltm	2

IPI00120095.2	R.DFTK@PQDGDIIAPLIT*PLK@.W	115	Samhd1	2

IPI00340860.5	R.TASEGSEAETPEAPK.Q	116	Larp7	2

IPI00664670.4	R.LGS*FGSITR{circumflex over ( )}.Q	117	Flnc	2

IPI00654192.2	R.S*LSFSEPQQPPPTVK@.S	118	Zfp395	2

IPI00225062.2	K.IHTTALTGQSPPLASGHQGEGDAPSVEPGATNIQQPSS	119	Srrm2	2
	PAPSTK.Q

IPI00225062.2	K.IHTTALTGQSPPLASGHQGEGDAPSVEPGATNIQQPSS	120	Srrm2	2
	PAPST*K@.Q

IPI00225062.2	K.IHTTALTGQSPPLASGHQGEGDAPSVEPGATNIQQPSS	121	Srrm2	2
	PAPST*K.Q

IPI00339428.9	R.MSSHTETS*SFLQTLTGR.L	122	Dock7	2

IPI00225062.2	R.DGSGTPSRHSLSGSSPGMKDTPQT*PSR.G	123	Srrm2	2

IPI00668709.2	R.DRS*SSAPNVHINTIEPVNIDDLIR.D	124	Braf	2

IPI00135660.5	R.RGNNSAVGSNADLT*IEEDEEEEPVALQQAQQVR.Y	125	Sdpr	2

IPI00135660.5	R.R{circumflex over ( )}GNNSAVGSNADLTIEEDEEEEPVALQQAQQVR{circumflex over ( )}.Y	126	Sdpr	2

IPI00404545.2	R.SLSSPTVTLSAPLEGAK@.D	127	Nedd4l	2

IPI00121892.9	R.R{circumflex over ( )}PPS*PDPNTK@.V	128	Spnb2	2

IPI00322707.5	K.AKQPVIGDQNSDS*DEMLAVLK.E	129	Atrx	2

IPI00929786.1	R.SLPTTVPESPNYR{circumflex over ( )}.N	130	Larp1	3

IPI00125319.1	R.TTS*FAESCK@PVQQPSAFGSMK@.V	131	Gsk3b	3

Table 3. Categorization of mTOR targets identified in the rapamycin and Ku-0063794 screens. Class 1 includes downstream effectors of rapamycin-sensitive mTORC1. Class 2 includes downstream effectors of rapamycin-insensitive mTORC1 or mTORC2. Class 3 includes the proteins downstream of both mTORC1 and mTORC2. Name of exemplary genes and sequences of respective phosphopeptides are shown.
*represents the site of phosphorylation (Ser, Thr and Tyr);
{circumflex over ( )}and @ represent heavy Arg and Lys, respectively.

TABLE 4

Extracted Term	IPI References

Rapamycin screen

mTOR signaling pathway	IPI00453603, IPI00318938, IPI00399440, IPI00268673, IPI00467843, IPI00221581, IPI00121335
ErbB signaling pathway	IPI00125319, IPI00453603, IPI00318938, IPI00113563, IPI00268673, IPI00121335
Insulin signaling pathway	IPI00125319, IPI00453603, IPI00116923, IPI00318938, IPI00268673, IPI00379844, IPI00121335
Acute myeloid leukemia	IPI00453603, IPI00318938, IPI00268673, IPI00121335
Focal adhesion	IPI00403938, IPI00125319, IPI00131138, IPI00113563, IPI00671847, IPI00121335
Tight junction	IPI00118143, IPI00380354, IPI00323349, IPI00845596, IPI00121335
Pathways in cancer	IPI00125319, IPI00121418, IPI00381495, IPI00113563, IPI00268673, IPI00380817, IPI00121335
Prostate cancer	IPI00125319, IPI00121418, IPI00268673, IPI00121335
RNA degradation	IPI00169888, IPI00309059, IPI00330066
Glioma	IPI00121418, IPI00268673, IPI00121335
Adipocytokine signaling pathway	IPI00268673, IPI00379844, IPI00121335
VEGF signaling pathway	IPI00111169, IPI00113563, IPI00121335
Chronic myeloid leukemia	IPI00121418, IPI00380817, IPI00121335

Ku-0063794 Screen

Erb6 signaling pathway	IPI00453999, IPI00125319, IPI00453603, IPI00230719, IPI00318938, IPI00668709, IPI00131999
mTOR signaling pathway	IPI00453603, IPI00230719, IPI00318938, IPI00668709, IPI00221581
Acute myeloid leukemia	IPI00453603, IPI00230719, IPI00318938, IPI00668709, IPI00131999
Cell cycle	IPI00125319, IPI00124717, IPI00137864, IPI00131999
Insulin signaling pathway	IPI00125319, IPI00453603, IPI00230719, IPI00318938, IPI00668709
Endometrial cancer	IPI00125319, IPI00230719, IPI00668709, IPI00131999
MAPK signaling pathway	IPI00230719, IPI00117088, IPI00123474, IPI00668709, IPI00664670, IPI00131999
RNA degradation	IPI00309059, IPI00119442, IPI00330066
Focal adhesion	IPI00125319, IPI00230719, IPI00405227, IPI00668709, IPI00664670
Colorectal cancer	IPI00125319, IPI00230719, IPI00668709, IPI00131999
TGF-beta signaling pathway	IPI00453603, IPI00137864, IPI00131999

Table 4. Gene ontology analysis of mTOR targets identified in the rapamycin and Ku-0063794 screens. Pathways that were overrepresented among the rapamycin-sensitive and Ku-sensitive mTOR targets are shown. Also shown is the biological process that the rapamycin-sensitive and Ku-sensitive targets overrepresented.

TABLE 5

Summary of the data. Two biological replicates were obtained for the rapamycin
screen (each contained data from two technical replicates) and one SILAC experiment was
performed for the Ku-0063794 screen. (L), light cells. (H), heavy cells.

					# of	# of	# of		Total
	Bio-	# of tech-			phospho-	phospho-	unique	Total	phospho-
Screen	repli-	replicate	Cell	Drug	peptides	proteins	phospho-	unique	proteins
ID	cate	combined	line	treatment	(FDR %)	(FDR %)	sites	sites	(FDR %)

Rapa	#	1	2	TSC2-/-	(L) DMSO	14,635	1,615	4,484
				(H) rapa	(0.25%)	(1.0%)
	#2	2	TSC2-/-	(L) rapa	32,500	1,866	6,832	11,271	2,778
				(H) DMSO	(0.28%)	(0.97%)			(1.7%)
Ku	#	1	1	wt MEF	(L) rapa	34,642	1,867	6,220
				(H) rapa+Ku	(0.28%)	(0.97%)

TABLE 6

Gene			SEQ	Amino acid	Median
Name	Description	Modified Sequence	ID NO	position	(treated/ctrl)

Rapa Screen (Proteins with downregulated phosphorylation)

RPS6	Ribosomal protein S6	RLSSLRASTSK	132	S235, S236	30.5

RPS6	Ribosomal protein S6	LSSLRASTSKSESSQK	133	S240, S242,	4.5
				S246, S247

Akt1s1	Proline-rich AKT1 substrate 1	S*LPVSVPVWAFK	134	S184	2.2

Gsk3β	Glycogen synthase kinase-3	TTS*FAESCKPVQQPSAFGSMK	135	S9	3.3
	beta

Eif4ebp2	Eukaryotic translation initiation	RNSPMAQTPPCHLPNIPGVT	136	S65, T70	39.1
	factor 4E-binding protein 2	SPGALIEDSK

Eif4ebp2	Eukaryotic translation initiation	TVAISDAAQLPDYCTT*PGGT	137	T37, T46	2.3
	factor 4E-binding protein 2	LFSTT*PGGTRIIYDRK

Eif4b	Eukaryotic translation initiation	SRTGSESSQTGASATSGR	138	T420, S422	4.3
	factor 4B

Eif4b	Eukaryotic translation initiation	TGSESSQTGASATSGR	139	S422, S425	4.6
	factor 4B

Eif4ebp1	Eukaryotic translation initiation	NSPVAKTPPKDLPAIPGVTSP	140	S64, T69	2.8
	factor 4E-binding protein 1	TSDEPPMQASQSQLPSSPEDK

Eif4g1	Isoform 1 of Eukaryotic	SFS*KEVEER	141	S1189	2.8
	translation initiation factor 4
	gamma 1

Rps6kb1	Isoform Alpha I of Ribosomal	FIGSPRTPVS*PVKFSPGDFW	142	S441, T444,	2.8
	protein S6 kinase beta-1	GR		S447

mTOR	Isoform 1 of FKBP12-	AGTTVPESIHSFIGDGLVKPE	143	S2478,	3.6
	rapamycin complex-	ALNKK		S2481
	associated protein

ULK1	Serine/threonine-protein	GGGASSPAPVVFTVGSPPSG	144	S747, S757,	4.1
	kinase ULK1	AT*PPQSTR		T763

Grb10	Isoform 3 of Growth factor	MNILSSQSPLHPSTLNAVIHR	145	S421, S423	27.3
	receptor-bound protein 10

Grb10	Isoform 3 of Growth factor	TASLPAIPNPFPELTGAAPGS	146	T76, S96,	5.4
	receptor-bound protein 10	PPSVAPSS*LPPPPSQPPAK		S104

Ku Screen (Proteins with downregulated phosphorylation)

Flnc	Filamin-C	LGS*FGSITR	147	S2234	2.4

Gsk3β	Glycogen synthase kinase-3	TTS*FAESCKPVQQPSAFGSMK	148	S9	2.2
	beta

Ndrg1	Protein NDRG1	TASGSSVTSLEGTR	149	S330, S333	4.1

Braf	B-Raf protein	DRSSS*APNVHINTIEPVNIDDLIR	150	S312	2.1

Akt1s1	Proline-rich AKT1 substrate 1	LNT*SDFQK	151	T247	31.3

Eif4b	Eukaryotic translation initiation	TGS*ESSQTGASATSGR	152	S422	2.2
	factor 4B

Eif4ebp1	Eukaryotic translation initiation	VALGDGVQLPPGDYSTT*PGGT	153	T36, T45	7.2
	factor 4E-binding protein 1	LFSTT*PGGTR

Eif4ebp2	Eukaryotic translation initiation	NS*PMAQTPPCHLPNIPGVTSP	154	S65	6.8
	factor 4E-bindg protein 2	GALIEDSK

Table 6. Representative hits identified in the rapamycin and Ku-0063794 screens. Only protein targets with downregulated phosphorylation are shown. Median fold-changes of the identified phosphopeptides are reported. For the rapamycin screen, hits from the second biological replicate are shown. The control group is DMSO vehicle treated sample whereas the experiment group is the one treated with rapamycin. For the Ku-0063794 screen, the control group was treated with rapamycin whereas the experiment group was treated with a combination of rapamycin and Ku-0063794. An asterisk indicates the site of phosphorylation. Phosphorylation of Grb10 in MNILSSQSPLHPSTLNAVIHR (SEQ ID NO: 27) corresponds to S421/S423 in isoform 3 and S501/S503 in isoform 1, respectively. The site designation in the text corresponds to the sites in isoform 1 (mouse sequence).

TABLE 7

					Log2
		SEQ			Area
		ID	Gene		(H/L)
Reference	Ascore Seq	NO:	Name	Annotation	Ratio

Rapa replicate 1 (light cells were control and heavy cells were treated

with rapamycin. L, light; H, heavy)

IPI:IPI00136419.1	LDGNPINLS*R	155	Epyc	Epiphycan	−7.993

IPI:IPI00108454.2	LSSLRASTSKSESSQK	156	rpS6	29 kDa protein	−7.693

IPI:IPI00108454.2	RLSSLRASTSK	157	rpS6	29 kDa protein	−6.817

IPI:IPI00225062.2	TPQAPTPANLVVGPR{circumflex over ( )}S*A	158	Srrm2	Isoform 3 of Serine/arginine repetitive	−6.606
	HGTAPVNIAGSR{circumflex over ( )}			matrix protein 2

IPI:IPI00111497.1	VTEGSYSCLAHS*PLGVVA	159	Igdcc4	Isoform 2 of Immunoglobulin	−6.062
	SQVAVVK			superfamily DCC subclass member 4

IPI:IPI00874813.1	MPAT*DTANLTAPWHPR	160	Ldhd	22 kDa protein	−5.745

IPI:IPI00126317.1	NASTSFQELEDKK	161	Dnajc2	DnaJ homolog subfamily C member 2	−5.522

IPI:IPI00187443.1	WLKEAEEESSGGEEEDE	162	Eif5;	Eukaryotic translation initiation	−5.378
	DENIEVVYSK		LOC100047658	factor 5

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	163	Eif4ebp1	Eukaryotic translation initiation	−5.301
	PGGTLFS*TTPGGTR			factor 4E-binding protein 1

IPI:IPI00225062.2	SAHGTAPVNIAGSRTPAG	164	Srrm2	Isoform 3 of Serine/arginine repetitive	−5.098
	LAPTNLS*SSR			matrix protein 2

IPI:IPI00467423.3	GYS*YDDSMESR	165	Gm13697	Novel protein containing MIF4G and	−5.093
				MA3 domains

IPI:IPI00467423.3	GYS*YDDSMESR	166	Gm13697	Novel protein containing MIF4G and	−4.733
				MA3 domains

IPI:IPI00130920.1	SDISPLT*PR	167	Mtap1b	Microtubule-associated protein 1B	−4.668

IPI:IPI00133685.1	AATATRPPGPPPAPQPPS*	168	Akt1s1	Proline-rich AKT1 substrate 1	−4.667
	PAPS*PPPRPALAR

IPI:IPI00154084.2	GFGDS*FGR	169	Myef2	Isoform 2 of Myelin expression factor 2	−4.553

IPI:IPI00338745.4	QADVADQQTTELPAENGET	170	Hmgn1;	Non-histone chromosomal protein	−4.546
	ENQSPASEEEKEAKSD		LOC100044391	HMG-14

IPI:IPI00109318.1	RNSPMAQTPPCHLPNIPG	171	Eif4ebp2	Eukaryotic translation initiation	−4.531
	VTSPGALIEDSK			factor 4E-binding protein 2

IPI:IPI00154084.2	GFGDS*FGR	172	Myef2	Isoform 2 of Myelin expression factor 2	−4.429

IPI:IPI00121519.1	SYSESGLMGEPQPQGPPS*	173	Neurod1	Neurogenic differentiation factor 1	−4.393
	WTDECLSSQDEEHEADKK

IPI:IPI00317794.5	KEDSDEDEDEEDEDDSD	174	Ncl	Nucleolin	−4.345
	EDEDDEEEDEFEPPIVK

IPI:IPI00399953.1	VFTSDISDPVVASTSQAPG	175	Wnk1	Serine/threonine-protein kinase WNK1	−4.306
	M#NLSHSASSLS*LQQAFS
	ELK

IPI:IPI00263048.1	VS*SETHQGPGTPESK	176	Numa1	Nuclear mitotic apparatus protein 1	−4.079

IPI:IPI00225062.2	TPPSAPSQSRM#TSER	177	Srrm2	Isoform 3 of Serine/arginine repetitive	−4.002
				matrix protein 2

IPI:IPI00263048.1	VSS*ETHQGPGTPESK	178	Numa1	Nuclear mitotic apparatus protein 1	−3.674

IPI:IPI00664808.2	SSSPT*NSLTQPIEMPTLSS	179	Kdm3b	Isoform 2 of Lysine-specific	−3.64
	S*PTEER{circumflex over ( )}PTVGPGQQDNP			demethylase 3B
	LLK@

IPI:IPI00225062.2	RKETPS*PR	180	Srrm2	Isoform 3 of Serine/arginine repetitive	−3.569
				matrix protein 2

IPI:IPI00453603.1	TPVSPVKFSPGDFWGR	181	Rps6kb1	Isoform Alpha I of Ribosomal protein	−3.424
				S6 kinase beta-1

IPI:IPI00318938.6	R{circumflex over ( )}VALGDGVQLPPGDY*ST	182	Eif4ebp1	Eukaryotic translation initiation	−3.386
	TPGGTLFSTTPGGT*R{circumflex over ( )}			factor 4E-binding protein 1

IPI:IPI00330066.5	DSQDTSAEQS*DHDDEVAS	183	Edc4	Isoform 1 of Enhancer of mRNA-	−3.347
	LASASGGFGSK			decapping protein 4

IPI:IPI00399953.1	DAMNLS*GR	184	Wnk1	Serine/threonine-protein kinase WNK1	−3.343

IPI:IPI00127707.1	LHQLAMQQSHFPMTHGNT	185	Pcbp2	Isoform 1 of Poly(rC)-binding protein 2	−3.277
	GFSGIESSS*PEVK

IPI:IPI00458068.6	SNS*LSEQLTVNSNPDTVK	186	Fkbp15	Isoform B of FK506-binding protein 15	−3.252

IPI:IPI00116442.1	S*EGLSLER	187	Hdgfrp2	Isoform 3 of Hepatoma-derived growth	−3.127
				factor-related protein 2

IPI:IPI00116442.1	S*EGLSLER{circumflex over ( )}	188	Hdgfrp2	Isoform 3 of Hepatoma-derived growth	−3.127
				factor-related protein 2

IPI:IPI00112101.1	TTQSLQDFPVADS*EEEAE	189	Tfip11	Tuftelin-interacting protein 11	−3.096
	EEFQKELSQWR

IPI:IPI00750472.3	TESASLSQS*QVNGFFASH	190	Ptpn13	Tyrosine-protein phosphatase non-	−3.058
	LGDR			receptor type 13

IPI:IPI00318938.6	VALGDGVQLPPGDYS*TTP	191	Eif4ebp1	Eukaryotic translation initiation	−3.042
	GGTLFSTTPGGTR			factor 4E-binding protein 1

IPI:IPI00317599.3	SQEDEEEISTS*PGVSEFVS	192	Syap1	Synapse-associated protein 1	−3.022
	DAFDTCS*LNQEDLRK

IPI:IPI00116442.1	S*EGLSLER	193	Hdgfrp2	Isoform 3 of Hepatoma-derived growth	−3.012
				factor-related protein 2

IPI:IPI00553798.2	TPEMIIQKPKISM#QDVDL	194	Ahnak	AHNAK nucleoprotein isoform 1	−2.987
	S*LGSCK

IPI:IPI00751009.1	TSEFPTPLFSGPLEPVACG	195	Srpk2	serine/arginine-rich protein specific	−2.951
	SVISEGSPLTEQEESSPSH			kinase 2
	DR{circumflex over ( )}S*R{circumflex over ( )}

IPI:IPI00127976.1	MNILSSQS*PLHPSTLNAVI	196	Grb10	Isoform 3 of Growth factor receptor-	−2.946
	HR			bound protein 10

IPI:IPI00127976.1	M#NILSS*QSPLHPSTLNAVI	197	Grb10	Isoform 3 of Growth factor receptor-	−2.944
	HR			bound protein 10

IPI:IPI00656285.2	SSGLQTPECLSREGSPIP	198	Foxk1	Forkhead box protein K1	−2.896
	HDPDLGSK

IPI:IPI00830829.1	SGEQITSSPVS*PK	199	Hisppd1	140 kDa protein	−2.883

IPI:IPI00380722.1	LAWVGDVFTTT*PTDPRPL	200	Bat2	Large proline-rich protein BAT2	−2.824
	TS*PLR

IPI:IPI00153986.2	GTS*RPGTPSAEAASTSSTLR	201	Gtf2f1	General transcription factor IIF	−2.802
				subunit 1

IPI:IPI00129276.2	LES*LNIQR{circumflex over ( )}	202	Eif3a	Eukaryotic translation initiation	−2.792
				factor 3 subunit A

IPI:IPI00121277.1	SAS*ESYTQSFQSR	203	Pi4k2a	Phosphatidylinositol 4-kinase type 2-	−2.789
				alpha

IPI:IPI00470003.3	SPTPKSPPSR	204	Parva	Alpha-parvin	−2.746

IPI:IPI00308971.1	AEALSSLHGDDQDS*EDEV	205	Igf2r	Cation-independent mannose-6-	−2.738
	LTVPEVK			phosphate receptor

IPI:IPI00121277.1	SASESYT*QSFQSR	206	Pi4k2a	Phosphatidylinositol 4-kinase type 2-	−2.723
				alpha

IPI:IPI00130920.1	VLSPLR{circumflex over ( )}SPPLLGSESPYE	207	Mtap1b	Microtubule-associated protein 1B	−2.715
	DFLSADSK@

IPI:IPI00129276.2	LES*LNIQR	208	Eif3a	Eukaryotic translation initiation	−2.711
				factor 3 subunit A

IPI:IPI00111169.1	CSVS*LSNVEAR	209	Pla2g4a	Cytosolic phospholipase A2	−2.664

IPI:IPI00458068.6	HSSGNSM#LLPSMSVTM	210	Fkbp15	Isoform B of FK506-binding protein 15	−2.641
	ETSM#IM#SNIQR

IPI:IPI00751009.1	TVSASSTGDLPK@	211	Srpk2	serine/arginine-rich protein-specific	−2.607
				kinase 2

IPI:IPI00222828.2	S*SPNPFVGSPPK@	212	Dab2	Isoform p93 of Disabled homolog 2	−2.599

IPI:IPI00111169.1	CSVS*LSNVEAR	213	Pla2g4a	Cytosolic phospholipase A2	−2.579

IPI:IPI00120886.3	SVGDGET*VEFDVVEGEK	214	Ybx1	Nuclease-sensitive element-binding	−2.553
				protein 1

IPI:IPI00751009.1	TVSASSTGDLPK@	215	Srpk2	serine/arginine-rich protein-specific	−2.534
				kinase 2

IPI:IPI00133349.1	DR{circumflex over ( )}SPSPLR{circumflex over ( )}GNVVPS*PL	216	Carhsp1	Calcium-regulated heat stable protein 1	−2.512
	PTR{circumflex over ( )}

IPI:IPI00469392.2	GSGSVDET*LFALPAASEPV	217	Rtn4	Isoform 1 of Reticulon-4	−2.475
	IPSSAEK

IPI:IPI00127976.1	T*ASLPAIPNPFPELTGAAP	218	Grb10	Isoform 3 of Growth factor receptor-	−2.437
	GSPPSVAPSSLPPPPSQP			bound protein 10
	PAK

IPI:IPI00751009.1	TSEFPTPLFSGPLEPVACG	219	Srpk2	serine/arginine-rich protein-specific	−2.408
	SVISEGSPLTEQEESSPSH			kinase 2
	DRS*R

IPI:IPI00761677.1	S*RSPVDSPVPASMFAPEP	220	Gtpbp1	GTP-binding protein 1	−2.406
	SS*PGAAR

IPI:IPI00468516.3	TRTVLSLFDEDEDKVEDE	221	D6Wsu116e	Isoform 1 of Protein FAM21	−2.398
	SSTCAPQDGR

IPI:IPI00396728.3	R{circumflex over ( )}VSGS*ATPNSEAPR{circumflex over ( )}	222	Ddx51	ATP-dependent RNA helicase DDX51	−2.351

IPI:IPI00660767.2	KRSDSSGGYTLSDVIQS*	223	Ibtk	Isoform 2 of Inhibitor of Bruton	−2.347
	PPSAGLLK			tyrosine kinase

IPI:IPI00226750.2	TSSTCSNES*LNAGGTPV	224	Tbc1d4	140 kDa protein	−2.275
	TPR

IPI:IPI00113033.1	RSPASSGSVAPQPS*SPPS	225	Ighmbp2	DNA-binding protein SMUBP-2	−2.241
	*PAQAEPEPR

IPI:IPI00136107.1	THSTSSSIGSGESPFSR	226	Ndrg3	Protein NDRG3	−2.202

IPI:IPI00751009.1	TSEFPTPLFSGPLEPVACG	227	Srpk2	serine/arginine-rich protein-specific	−2.187
	SVISEGSPLTEQEESSPSH			kinase 2
	DRS*R

IPI:IPI00321647.2	QNPEQS*ADEDAEKNEEDS	228	Eif3c	Eukaryotic translation initiation	−2.165
	EGSS*DEDEDEDGVGNTT			factor 3 subunit C
	FLKK

IPI:IPI00453800.4	HSSFADTGAAPSALSPESP	229	Zfp828	Zinc finger protein 828	−2.147
	VLATSPEPWGPSLSASPE
	SR{circumflex over ( )}

IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	230	Ndrg3	Protein NDRG3	−2.124

IPI:IPI00377615.2	RSSRSFSLDEPPLFIPDNI	231	Phf3	PHD finger protein 3	−2.124
	ATVK

IPI:IPI00377615.2	R{circumflex over ( )}SSR{circumflex over ( )}SFSLDEPPLFIPD	232	Phf3	PHD finger protein 3	−2.124
	NIATVK@

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	233	Npm1	Nucleophosmin	−2.101
	AES*EDEDEEDVKLLGM#S
	GK

IPI:IPI00331075.4	SLNFQEDDDTSRETFASD	234	Phax	Phosphorylated adapter RNA export	−2.101
	TNEALASLDEAQEGPGETK			protein

IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	235	Ndrg3	Protein NDRG3	−2.098

IPI:IPI00136107.1	THSTSSSIGSGESPFSR	236	Ndrg3	Protein NDRG3	−2.098

IPI:IPI00669522.4	SRS*DIDVNAAASAK	237	Clasp1	Isoform 2 of CLIP-associating protein 1	−2.073

IPI:IPI00117953.2	STDSPVAGSEDDLVAAAP	238	Bicc1;	Isoform 1 of Protein bicaudal C	−2.07
	LLHSPEWSEER{circumflex over ( )}		4930533K18Rik	homolog 1

IPI:IPI00137229.1	GFQFVSSS*LPDICYR	239	Cnbp	Isoform 2 of Cellular nucleic acid-	−2.061
				binding protein

IPI:IPI00380309.3	AMLDQLMGTS*R	240	Luc7l2	Isoform 1 of Putative RNA-binding	−2.046
				protein Luc7-like 2

IPI:IPI00153986.2	GTSRPGT*PSAEAASTSSTLR	241	Gtf2f1	General transcription factor IIF	−2.035
				subunit 1

IPI:IPI00330262.5	S*QDDAMVDYFFQR	242	Pum1	Isoform 1 of Pumilio homolog 1	−2.02

IPI:IPI00312128.3	LDLDLTS*DSQPPVFK	243	Trim28	Isoform 1 of Transcription intermediary	−2.01
				factor 1-beta

IPI:IPI00667973.3	IRQPSIDLTDDDQTSSVPH	244	Cep170	Isoform 1 of Centrosomal protein	−1.979
	SAISDIMSSDQETYSCK			of 170 kDa

IPI:IPI00468516.3	TRTVLSLFDEDEDKVEDE	245	D6Wsu116e	Isoform 1 of Protein FAM21	−1.956
	SSTCAPQDGR

IPI:IPI00136207.6	YTPTSPSY*SPSSPEYTPAS	246	Polr2a	DNA-directed RNA polymerase II	−1.95
	PK@			subunit RPB1

IPI:IPI00225062.2	SLLPNSSQDELM#EVEKSE	247	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.937
	QPLSQVLPSLS*PEHK			matrix protein 2

IPI:IPI00222828.2	S*SPNPFVGSPPK@	248	Dab2	Isoform p93 of Disabled homolog 2	−1.915

IPI:IPI00309059.7	RSTSPIIGS*PPVR	249	Patl1	Protein PAT1 homolog 1	−1.901

IPI:IPI00309059.7	RSTSPIIGS*PPVR	250	Patl1	Protein PAT1 homolog 1	−1.896

IPI:IPI00338745.4	QADVADQQTTELPAENGET	251	Hmgn1;	Non-histone chromosomal protein	−1.895
	ENQSPASEEEKEAKS*D		LOC100044391	HMG-14

IPI:IPI00417158.1	SQTPQLHFLDT*DDEISPTS	252	Smg6	Telomerase-binding protein EST1A	−1.895
	WGDSR{circumflex over ( )}

IPI:IPI00761759.1	SSS*GSEHSTEGSVSLGDG	253	Larp4	Putative uncharacterized protein	−1.89
	PLSR

IPI:IPI00314502.5	VHGLPT*TSPSGVNMAELA	254	Tcfeb	Transcription factor EB	−1.887
	QQVVK

IPI:IPI00117689.1	ATEEPSGTGS*DELIK@SD	255	Ptrf	Polymerase I and transcript release	−1.875
	QVNGVLVLSLLDK@			factor

IPI:IPI00462949.4	AENIMAS*LER	256	Dock11	Dedicator of cytokinesis protein 11	−1.851

IPI:IPI00471475.1	FDQLFDDES*DPFEVLK@	257	Serbp1	Isoform 1 of Plasminogen activator	−1.845
				inhibitor 1 RNA-binding protein

IPI:IPI00471475.1	FDQLFDDES*DPFEVLK	258	Serbp1	Isoform 1 of Plasminogen activator	−1.845
				inhibitor 1 RNA-binding protein

IPI:IPI00309059.7	RSTSPIIGS*PPVR	259	Patl1	Protein PAT1 homolog 1	−1.837

IPI:IPI00331612.3	K@PAQETEETSSQESAE	260	Hmga2	High mobility group protein HMGI-C	−1.834
	ED

IPI:IPI00929786.1	SLPTTVPESPNYR{circumflex over ( )}	261	Larp1	Isoform 1 of La-related protein 1	−1.823

IPI:IPI00112101.1	TTQSLQDFPVADS*EEEAE	262	Tfip11	Tuftelin-interacting protein 11	−1.813
	EEFQKELSQWR

IPI:IPI00133685.1	SSDEENGPPSS*PDLDR	263	Akt1s1	Proline-rich AKT1 substrate 1	−1.779

IPI:IPI00761443.2	SLS*ESYELSSDLQDK	264	Iqsec1	IQ motif and Sec7 domain 1 isoform b	−1.779

IPI:IPI00318938.6	NS*PVAKTPPKDLPAIPGVT	265	Eif4ebp1	Eukaryotic translation initiation	−1.776
	*SPTSDEPPMQASQSQLPS			factor 4E-binding protein 1
	SPEDK

IPI:IPI00318938.6	NSPVAKT*PPKDLPAIPGVT	266	Eif4ebp1	Eukaryotic translation initiation	−1.776
	SPT*SDEPPMQASQSQLPS			factor 4E-binding protein 1
	SPEDK

IPI:IPI00421179.1	SFS*KEVEER	267	Eif4g1	Isoform 1 of Eukaryotic translation	−1.77
				initiation factor 4 gamma 1

IPI:IPI00379844.4	TYS*LTTPAR	268	Irs2	Insulin receptor substrate 2	−1.769

IPI:IPI00379844.4	TYS*LTTPAR	269	Irs2	Insulin receptor substrate 2	−1.769

IPI:IPI00127989.1	FSEM#M#DHMGGDEDVDL	270	Ptges3;	Prostaglandin E synthase 3	−1.762
	PEVDGADDDSQDSDDEK		Gm9769;
	MPDLE		LOC100048119

IPI:IPI00127989.1	FSEM#M#DHMGGDEDVDL	271	Ptges3;	Prostaglandin E synthase 3	−1.762
	PEVDGADDDSQDSDDEK		Gm9769;
	@MPDLE		LOC100048119

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAK	272	Eef1b2	Elongation factor 1-beta	−1.754
	@DDDDIDLFGS*DDEEESE
	EAK@

IPI:IPI00318938.6	NSPVAK@TPPK@DLPAI	273	Eif4ebp1	Eukaryotic translation initiation	−1.752
	PGVTSPTSDEPPMQASQS			factor 4E-binding protein 1
	QLPSSPEDK@

IPI:IPI00317794.5	KEDSDEDEDEEDEDDSD	274	Ncl	Nucleolin	−1.75
	EDEDDEEEDEFEPPIVK

IPI:IPI00116331.1	SRTPSASHEEQQE	275	Sgta	Isoform 1 of Small glutamine-rich	−1.747
				tetratricopeptide repeat-containing
				protein alpha

IPI:IPI00116331.1	SR{circumflex over ( )}TPSASHEEQQE	276	Sgta	Isoform 1 of Small glutamine-rich	−1.747
				tetratricopeptide repeat-containing
				protein alpha

IPI:IPI00553798.2	FGTFGGLGSK	277	Ahnak	AHNAK nucleoprotein isoform 1	−1.745

IPI:IPI00130510.1	LQDISELLATGVGLSDSEV	278	Ppan	Suppressor of SWI4 1 homolog	−1.743
	EPDGEHNTTELPQAVAGR{circumflex over ( )}

IPI:IPI00330262.5	S*QDDAMVDYFFQR	279	Pum1	Isoform 1 of Pumilio homolog 1	−1.734

IPI:IPI00459468.2	LSESSALKQPATPTAAES	280	Wmip1	Isoform 1 of ATPase WRNIP1	−1.728
	S*EGEGEEGDDGGETESR

IPI:IPI00130573.1	SLLSHEFQDETDTEEETL	281	Cpd	Carboxypeptidase D	−1.723
	YSSK

IPI:IPI00468516.3	TSALLFSS*DEEDQWNIADS	282	D6Wsu116e	Isoform 1 of Protein FAM21	−1.718
	HTK

IPI:IPI00468516.3	TSALLFSS*DEEDQWNIADS	283	D6Wsu116e	Isoform 1 of Protein FAM21	−1.718
	HTK

IPI:IPI00109676.6	SSGSSSSGLGTVSS*SPAS	284	Phf8	Isoform 1 of PHD finger protein 8	−1.715
	QR

IPI:IPI00399953.1	VFTSDISDPVVAST*SQAPG	285	Wnk1	Serine/threonine-protein kinase WNK1	−1.704
	MNLSHSASSLSLQQAFSELK

IPI:IPI00399953.1	VFTSDISDPVVASTSQAPG	286	Wnk1	Serine/threonine-protein kinase WNK1	−1.704
	MNLSHSASS*LSLQQAFSE
	LK@

IPI:IPI00225062.2	RSSSELSPEVVEK	287	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.702
				matrix protein 2

IPI:IPI00116331.1	SRTPSASHEEQQE	288	Sgta	Isoform 1 of Small glutamine-rich	−1.694
				tetratricopeptide repeat-containing
				protein alpha

IPI:IPI00116331.1	SRTPSASHEEQQE	289	Sgta	Isoform 1 of Small glutamine-rich	−1.69
				tetratricopeptide repeat-containing
				protein alpha

IPI:IPI00116331.1	SR{circumflex over ( )}TPSASHEEQQE	290	Sgta	Isoform 1 of Small glutamine-rich	−1.69
				tetratricopeptide repeat-containing
				protein alpha

IPI:IPI00405752.3	VLQESDVSPSSSTTSLPIS	291	C130092O11Rik	Isoform 1 of Uncharacterized protein	−1.689
	PLTEEPLPFKDITR			KIAA1680

IPI:IPI00380415.2	ALSLSSADS*TDAKR	292	Bat2l	Isoform 1 of Protein BAT2-like	−1.687

IPI:IPI00130510.1	LQDISELLATGVGLSDSEV	293	Ppan	Suppressor of SWI4 1 homolog	−1.68
	EPDGEHNTTELPQAVAGR

IPI:IPI00229859.1	AKPAAQSEEETATSPAAS	294	Eif3b	Eif3b protein	−1.677
	PTPQSAERSPSQEPSAP
	GK

IPI:IPI00224200.1	RLSSTGGQTPR	295	Iws1	Isoform 1 of Protein IWS1 homolog	−1.663

IPI:IPI00225062.2	R{circumflex over ( )}SSSELSPEVVEK@	296	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.663
				matrix protein 2

IPI:IPI00331612.3	K@PAQETEETSSQESAE	297	Hmga2	High mobility group protein HMGI-C	−1.658
	ED

IPI:IPI00229645.2	K@GSSGNASEVSVACLT	298	Cytsa	Cytospin-A	−1.657
	ER{circumflex over ( )}

IPI:IPI00317794.5	KEDSDEDEDEEDEDDSD	299	Ncl	Nucleolin	−1.637
	EDEDDEEEDEFEPPIVK

IPI:IPI00663627.1	GFLDGVYSFEYYPS*TPGK	300	Flnb	Filamin-B	−1.621

IPI:IPI00312128.3	LDLDLTSDS*QPPVFK	301	Trim28	Isoform 1 of Transcription intermediary	−1.616
				factor 1-beta

IPI:IPI00225062.2	HSGSTSPYPK@	302	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.603
				matrix protein 2

IPI:IPI00312414.4	TSTFCGT*PEFLAPEVLTET	303	Pkn2	Isoform 1 of Serine/threonine-protein	−1.603
	SYTR{circumflex over ( )}			kinase N2

IPI:IPI00553798.2	SNSFSDER{circumflex over ( )}EFSAPST*PT	304	Ahnak	AHNAK nucleoprotein isoform 1	−1.602
	GT*LEFAGGDAK@

IPI:IPI00126176.3	VSLLGPVTTPEFQLVKTPL	305	Racgap1	Rac GTPase-activating protein 1	−1.597
	SSSLSQR

IPI:IPI00225062.2	GSLSRSSSPVTELTAR	306	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.593
				matrix protein 2

IPI:IPI00399953.1	VFTSDISDPVVASTSQAPG	307	Wnk1	Serine/threonine-protein kinase WNK1	−1.588
	MNLSHS*ASSLSLQQAFSE
	LK

IPI:IPI00226155.3	VSALEEDMDDVESSEEEE	308	Prpf38a	Isoform 1 of Pre-mRNA-splicing factor	−1.585
	EEDEKLER			38A

IPI:IPI00226155.3	VSALEEDMDDVESSEEEE	309	Prpf38a	Isoform 1 of Pre-mRNA-splicing factor	−1.585
	EEDEK@LER{circumflex over ( )}			38A

IPI:IPI00116331.1	SRTPSASHEEQQE	310	Sgta	Isoform 1 of Small glutamine-rich	−1.58
				tetratricopeptide repeat-containing
				protein alpha

IPI:IPI00116331.1	SR{circumflex over ( )}TPSASHEEQQE	311	Sgta	Isoform 1 of Small glutamine-rich	−1.58
				tetratricopeptide repeat-containing
				protein alpha

IPI:IPI00135660.5	SLEETLHNVDLSSDDELP	312	Sdpr	Serum deprivation-response protein	−1.577
	R{circumflex over ( )}DEEALEDSAEEK@

IPI:IPI00323349.2	HSSHDMLSHS*WEGNR	313	Tjp2	Tight junction protein ZO-2	−1.566

IPI:IPI00169500.3	TESILDKEDKVPMAGVGGT	314	Atxn2l	Isoform 1 of Ataxin-2-like protein	−1.564
	EGPEQLPAPCPSQTGS*PP
	VGLIK

IPI:IPI00118143.1	K@QTPPASPS*PQPIEDR{circumflex over ( )}	315	Cttn	Src substrate cortactin	−1.556
	PPS*SPIYEDAAPFK@

IPI:IPI00127415.1	TVSLGAGAK@DELHIVEAE	316	Npm1	Nucleophosmin	−1.555
	AM#NYEGS*PIK@VTLATLK@

IPI:IPI00225062.2	SLLPNSSQDELMEVEK@S	317	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.55
	EQPLSQVLPSLS*PEHK@			matrix protein 2

IPI:IPI00225062.2	SLLPNSSQDELMEVEKSEQ	318	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.55
	PLSQVLPSLS*PEHK			matrix protein 2

IPI:IPI00320267.1	GDQVLNFS*DAEDLIDDSK	319	Mcrs1	Microspherule protein 1	−1.55
	@LK@

IPI:IPI00320267.1	GDQVLNFS*DAEDLIDDSKLK	320	Mcrs1	Microspherule protein 1	−1.55

IPI:IPI00553798.2	SNSFSDER{circumflex over ( )}EFSAPS*TPT	321	Ahnak	AHNAK nucleoprotein isoform 1	−1.548
	*GTLEFAGGDAK@

IPI:IPI00116331.1	SRTPSASHEEQQE	322	Sgta	Isoform 1 of Small glutamine-rich	−1.543
				tetratricopeptide repeat-containing
				protein alpha

IPI:IPI00222828.2	SSPNPFVGS*PPK	323	Dab2	Isoform p93 of Disabled homolog 2	−1.531

IPI:IPI00317794.5	KEDSDEDEDEEDEDDSD	324	Ncl	Nucleolin	−1.531
	EDEDDEEEDEFEPPIVK

IPI:IPI00225062.2	RSSSELSPEVVEK	325	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.524
				matrix protein 2

IPI:IPI00225062.2	R{circumflex over ( )}SSSELSPEVVEK@	326	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.524
				matrix protein 2

IPI:IPI00116331.1	SRTPSASHEEQQE	327	Sgta	Isoform 1 of Small glutamine-rich	−1.52
				tetratricopeptide repeat-containing
				protein alpha

IPI:IPI00380415.2	ALSLSS*ADSTDAKR	328	Bat2l	Isoform 1 of Protein BAT2-like	−1.519

IPI:IPI00225062.2	RSSSELSPEVVEK	329	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.518
				matrix protein 2

IPI:IPI00225062.2	R{circumflex over ( )}SSSELSPEVVEK@	330	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.518
				matrix protein 2

IPI:IPI00399469.2	VSPPASPTLDVELGAGEA	331	Sh3rf1	Isoform 2 of Putative E3 ubiquitin-	−1.504
	PLQGAVGPELPLGGSHGR			protein ligase SH3RF1

IPI:IPI00225062.2	R{circumflex over ( )}SSSELSPEVVEK@	332	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.501
				matrix protein 2

IPI:IPI00137501.1	TEMDKSPFNSPSPQDS*PR	333	Nfic	Isoform 1 of Nuclear factor 1 C-type	−1.498

IPI:IPI00309059.7	RSTSPIIGSPPVR	334	Patl1	Protein PAT1 homolog 1	−1.498

IPI:IPI00225062.2	RSSSELSPEVVEK	335	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.497
				matrix protein 2

IPI:IPI00116331.1	SR{circumflex over ( )}TPSASHEEQQE	336	Sgta	Isoform 1 of Small glutamine-rich	−1.493
				tetratricopeptide repeat-containing
				protein alpha

IPI:IPI00470095.1	HGSGADSDYENTQSGDP	337	Git1	ARF GTPase-activating protein GIT1	−1.488
	LLGLEGK@

IPI:IPI00470095.1	HGSGADSDYENTQSGDP	338	Git1	ARF GTPase-activating protein GIT1	−1.488
	LLGLEGK

IPI:IPI00225062.2	IHTTALTGQS*PPLASGHQG	339	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.479
	EGDAPSVEPGATNIQQPSS			matrix protein 2
	PAPSTK

IPI:IPI00116442.1	TRLAS*ESANDDNEDS	340	Hdgfrp2	Isoform 3 of Hepatoma-derived growth	−1.456
				factor-related protein 2

IPI:IPI00225062.2	GSLSRSSSPVTELTAR	341	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.454
				matrix protein 2

IPI:IPI00320267.1	GDQVLNFS*DAEDLIDDSK	342	Mcrs1	Microspherule protein 1	−1.454

IPI:IPI00225062.2	RSSSELSPEVVEK	343	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.445
				matrix protein 2

IPI:IPI00661508.3	S*IDPDSIQSALLASGLGSK@	344	Sbno1	Isoform 2 of Protein strawberry notch	−1.442
				homolog 1

IPI:IPI00661508.3	S*IDPDSIQSALLASGLGSK	345	Sbno1	Isoform 2 of Protein strawberry notch	−1.442
				homolog 1

IPI:IPI00317794.5	K@EDSDEDEDEEDEDDS	346	Ncl	Nucleolin	−1.441
	DEDEDDEEEDEFEPPIVK@

IPI:IPI00225267.6	ATPTESPEKPPPPAVQRDE	347	2310022B05Rik	Uncharacterized protein C1orf198	−1.438
	DDDALFS*EPALAQISSSNV			homolog
	LLK

IPI:IPI00122594.4	EVSVSSVTEEPKLDSSQL	348	Ahctf1	AT-hook-containing transcription	−1.437
	PLQTGLDVPATPR			factor 1

IPI:IPI00129356.1	SAFTPATATGSSPSPVLG	349	Itsn1	lsoform 1 of Intersectin-1	−1.43
	QGEK@VEGLQAQALYPWR{circumflex over ( )}

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGV	350	Eif4ebp1	Eukaryotic translation initiation	−1.412
	TSPTSDEPPMQASQSQLPS			factor 4E-binding protein 1
	S*PEDK

IPI:IPI00224200.1	VVCDADDS*DSDVVSDK@	351	Iws1	Isoform 1 of Protein IWS1 homolog	−1.403

IPI:IPI00321647.2	QNPEQSADEDAEK@NEED	352	Eif3c	Eukaryotic translation initiation	−1.399
	SEGSS*DEDEDEDGVGN			factor 3 subunit C
	TTFLK@

IPI:IPI00664808.2	VSESVADDSSSRDSFTQS	353	Kdm3b	Isoform 2 of Lysine-specific	−1.393
	LESLTSGLCK			demethylase 3B

IPI:IPI00122863.2	RGQS*PQPDQGP	354	Rarg	Isoform 2 of Retinoic acid receptor	−1.387
				gamma

IPI:IPI00453853.2	TPVASECTQTDGAEEVAGS	355	Bap1	Ubiquitin carboxyl-terminal hydrolase	−1.38
	CPQTTTHS*PPSK@			BAP1

IPI:IPI00132028.1	K@GSDDDGGDSPVQDID	356	Sap30	Histone deacetylase complex subunit	−1.376
	TPEVDLYQLQVNTLR{circumflex over ( )}			SAP30

IPI:IPI00132028.1	KGSDDDGGDSPVQDIDT	357	Sap30	Histone deacetylase complex subunit	−1.376
	PEVDLYQLQVNTLR			SAP30

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGV	358	Eif4ebp1	Eukaryotic translation initiation	−1.374
	TSPTSDEPPMQASQSQLPS			factor 4E-binding protein 1
	S*PEDK

IPI:IPI00116442.1	TRLAS*ESANDDNEDS	359	Hdgfrp2	Isoform 3 of Hepatoma-derived growth	−1.358
				factor-related protein 2

IPI:IPI00153986.2	GTSRPGTPSAEAASTSST	360	Gtf2f1	General transcription factor IIF	−1.348
	LR			subunit 1

IPI:IPI00226149.6	GSASES*PSASVAEATTTD	361	Rsl1d1	Putative uncharacterized protein	−1.342
	VQVTPTALLQLDR{circumflex over ( )}

IPI:IPI00226149.6	GS*ASESPSASVAEATTTD	362	Rsl1d1	Putative uncharacterized protein	−1.342
	VQVTPTALLQLDR{circumflex over ( )}

IPI:IPI00313307.3	SYQNSPSSEDGIRPLPEY	363	Med1	Isoform 4 of Mediator of RNA	−1.337
	STEK			polymerase II transcription subunit 1

IPI:IPI00656285.2	SLVSPIPSPTGTISVPNSC	364	Foxk1	Forkhead box protein K1	−1.327
	PAS*PR

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAK	365	Eef1b2	Elongation factor 1-beta	−1.324
	@DDDDIDLFGSDDEEES*E
	EAK@

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGV	366	Eif4ebp1	Eukaryotic translation initiation	−1.322
	TSPTSDEPPMQASQSQLPS			factor 4E-binding protein 1
	S*PEDK

IPI:IPI00136572.4	SSTHASVS*PASEPSR	367	Zc3hav1	Isoform 1 of Zinc finger CCCH-type	−1.318
				antiviral protein 1

IPI:IPI00116442.1	TRLASESANDDNEDS	368	Hdgfrp2	Isoform 3 of Hepatoma-derived growth	−1.315
				factor-related protein 2

IPI:IPI00116442.1	TR{circumflex over ( )}LASESANDDNEDS	369	Hdgfrp2	Isoform 3 of Hepatoma-derived growth	−1.315
				factor-related protein 2

IPI:IPI00226149.6	GS*ASESPSASVAEATTTD	370	Rsl1d1	Putative uncharacterized protein	−1.314
	VQVTPTALLQLDR{circumflex over ( )}

IPI:IPI00169500.3	R{circumflex over ( )}SPGGT*SPPNGGLPGPL	371	Atxn2l	Isoform 1 of Ataxin-2-like protein	−1.308
	TAT*AAPPGPPAAVSPCLG
	PAAAAGSGLR{circumflex over ( )}

IPI:IPI00467362.6	SESLIDASEDSQLEAAIR	372	Ubxn7	UBX domain-containing protein 7	−1.307

IPI:IPI00124753.3	RNTFIGTPY*WMAPEVIACD	373	Mink1	misshapen-like kinase 1 isoform 2	−1.302
	ENPDATYDYR

IPI:IPI00110247.7	SLSQS*FENLLDEPAYGLIQ	374	Tbc1d15	TBC1 domain family member 15	−1.301
	K@

IPI:IPI00276222.1	KLDDQS*EDEALELEGPLIM	375	Fermt2	Fermitin family homolog 2	−1.293
	PGSGSIYSS*PGLYSK

IPI:IPI00460668.2	EVDYSDS*LTEK@	376	Smarca4	Putative uncharacterized protein	−1.293

IPI:IPI00380736.1	STTPT*PSVITR	377	Zfp516	Zinc finger protein 516	−1.259

IPI:IPI00153986.2	GTSR{circumflex over ( )}PGTPSAEAASTSS	378	Gtf2f1	General transcription factor IIF	−1.251
	TLR{circumflex over ( )}			subunit 1

IPI:IPI00230486.3	LSSKLSAVSLR	379	Snx17	Sorting nexin-17	−1.24

IPI:IPI00116331.1	SR{circumflex over ( )}TPSASHEEQQE	380	Sgta	Isoform 1 of Small glutamine-rich	−1.239
				tetratricopeptide repeat-containing
				protein alpha

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGV	381	Eif4ebp1	Eukaryotic translation initiation	−1.238
	TSPTSDEPPMQASQSQLPS			factor 4E-binding protein 1
	SPEDK

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGV	382	Eif4ebp1	Eukaryotic translation initiation	−1.238
	TSPTSDEPPMQASQSQLPS			factor 4E-binding protein 1
	SPEDK

IPI:IPI00749954.8	LR{circumflex over ( )}TDNASDASESS*DAE	383	Jmjd1c	jumonji domain containing 1C	−1.235
	SSSK@

IPI:IPI00225062.2	SVSSPR{circumflex over ( )}T*K@	384	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.234
				matrix protein 2

IPI:IPI00136107.1	THS*TSSSIGSGESPFSRA	385	Ndrg3	Protein NDRG3	−1.23

IPI:IPI00136107.1	THST*SSSIGSGESPFSR	386	Ndrg3	Protein NDRG3	−1.23

IPI:IPI00113377.1	K@EESEES*EDDMGFGLFD	387	Rplp1	60S acidic ribosomal protein P1	−1.229

IPI:IPI00553798.2	GGVTGSPEASIS*GSK@G	388	Ahnak	AHNAK nucleoprotein isoform 1	−1.228
	DLK@

IPI:IPI00330240.7	S*QDAESYQNVVELK@	389	Peg3	Isoform 1 of Paternally-expressed gene	−1.221
				3 protein

IPI:IPI00122845.1	LGS*QEPER{circumflex over ( )}	390			−1.219

IPI:IPI00224200.1	VVCDADDS*DSDVVSDK@	391	Iws1	Isoform 1 of Protein IWS1 homolog	−1.219

IPI:IPI00465879.2	LCS*SSSSDTSPR{circumflex over ( )}	392	Zc3hc1	Isoform 1 of Nuclear-interacting	−1.207
				partner of ALK

IPI:IPI00116442.1	TR{circumflex over ( )}LASESANDDNEDS	393	Hdgfrp2	Isoform 3 of Hepatoma-derived growth	−1.206
				factor-related protein 2

IPI:IPI00116442.1	TRLASESANDDNEDS	394	Hdgfrp2	Isoform 3 of Hepatoma-derived growth	−1.206
				factor-related protein 2

IPI:IPI00322095.4	IDEPNT*PYHNMIGDDEDAY	395	Ppp1r2	Putative uncharacterized protein	−1.204
	S*DSEGNEVMTPDILAK

IPI:IPI00322095.4	IDEPNTPY*HNMIGDDEDAY	396	Ppp1r2	Putative uncharacterized protein	−1.204
	S*DSEGNEVMTPDILAK

IPI:IPI00387384.3	LLCQEETPEAT*EDERDEER	397	Mett10d	Isoform 1 of Putative methyltransferase	−1.202
				METT10D

IPI:IPI00330246.2	WAAHGTS*PEDFSLTLGAR{circumflex over ( )}	398	Phldb1	Isoform 2 of Pleckstrin homology-like	−1.202
				domain family B member 1

IPI:IPI00330246.2	WAAHGTS*PEDFSLTLGAR	399	Phldb1	Isoform 2 of Pleckstrin homology-like	−1.202
				domain family B member 1

IPI:IPI00678133.3	SSHQDATDPEALWGVHQT	400	Inf2	Isoform 1 of Inverted formin-2	−1.199
	EADSTS*EGPEDEAQR{circumflex over ( )}

IPI:IPI00465879.2	LCS*SSSSDTSPR{circumflex over ( )}	401	Zc3hc1	Isoform 1 of Nuclear-interacting	−1.189
				partner of ALK

IPI:IPI00313513.6	S*VDIFGSTPFQPFSVSASK	402	Bmp2k	Isoform 1 of BMP-2-inducible protein	−1.18
				kinase

IPI:IPI00465879.2	LCS*SSSSDTSPR	403	Zc3hc1	Isoform 1 of Nuclear-interacting	−1.172
				partner of ALK

IPI:IPI00465879.2	LCS*SSSSDTSPR	404	Zc3hc1	Isoform 1 of Nuclear-interacting	−1.171
				partner of ALK

IPI:IPI00553798.2	EFSAPSTPTGTLEFAGGD	405	Ahnak	AHNAK nucleoprotein isoform 1	−1.169
	AK@

IPI:IPI00317794.5	K@EDSDEDEDEEDEDDS	406	Ncl	Nucleolin	−1.167
	DEDEDDEEEDEFEPPIVK@

IPI:IPI00465879.2	LCS*SSSSDTSPR{circumflex over ( )}	407	Zc3hc1	Isoform 1 of Nuclear-interacting	−1.163
				partner of ALK

IPI:IPI00465879.2	LCS*SSSSDTSPR	408	Zc3hc1	Isoform 1 of Nuclear-interacting	−1.163
				partner of ALK

IPI:IPI00229739.4	LVVSSPTSPKGK	409	Plekhm1	Pleckstrin homology domain-containing	−1.159
				family M member 1

IPI:IPI00465879.2	LCS*SSSSDTSPR	410	Zc3hc1	Isoform 1 of Nuclear-interacting	−1.156
				partner of ALK

IPI:IPI00465879.2	LCS*SSSSDTSPR{circumflex over ( )}	411	Zc3hc1	Isoform 1 of Nuclear-interacting	−1.155
				partner of ALK

IPI:IPI00225616.3	DGAGSPLR{circumflex over ( )}TSPPSTPSS	412	Dennd2a	DENN domain-containing protein 2A	−1.15
	PDDTFFNLGDLQNGR{circumflex over ( )}

IPI:IPI00720110.2	VLTANSNPSS*PSAAK	413	1600027N09Rik	RIKEN cDNA 1600027N09 gene	−1.148

IPI:IPI00465879.2	LCS*SSSSDTSPR	414	Zc3hc1	Isoform 1 of Nuclear-interacting	−1.143
				partner of ALK

IPI:IPI00461396.6	STRHSPSALQDVFVELLS	415	Baz1a	bromodomain adjacent to zinc finger	−1.134
	PHSK			domain 1A

IPI:IPI00222828.2	SSPNPFVGS*PPK@	416	Dab2	Isoform p93 of Disabled homolog 2	−1.134

IPI:IPI00222828.2	SSPNPFVGS*PPK	417	Dab2	Isoform p93 of Disabled homolog 2	−1.134

IPI:IPI00222828.2	SSPNPFVGS*PPK	418	Dab2	Isoform p93 of Disabled homolog 2	−1.134

IPI:IPI00225062.2	GCSPPKSPEKPPQSTSS*	419	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.133
	ESCPPSPQPTK			matrix protein 2

IPI:IPI00553798.2	GGVTGSPEASIS*GSK@G	420	Ahnak	AHNAK nucleoprotein isoform 1	−1.132
	DLK@

IPI:IPI00318671.2	DK@PVYDELFYTLS*PINGK@	421	Ehd4	EH-domain containing 4-KJR	−1.131
				(Fragment)

IPI:IPI00129388.1	HDHAVSLLTAASPT*ISLL	422	Scrib	Isoform 3 of Protein LAP4	−1.13
	LER

IPI:IPI00336973.2	GLNLDGTPALSTLGGFS*PA	423	Ccnl1	Isoform 1 of Cyclin-L1	−1.124
	SKPSS*PR

IPI:IPI00336973.2	GLNLDGTPALSTLGGFS*PA	424	Ccnl1	Isoform 1 of Cyclin-L1	−1.124
	SK@PSS*PR{circumflex over ( )}

IPI:IPI00317401.6	METVSNASSSSNPSS*PGR{circumflex over ( )}	425	Pds5b	Isoform 1 of Sister chromatid cohesion	−1.12
				protein PDS5 homolog B

IPI:IPI00317401.6	METVSNASSSSNPSS*PGR	426	Pds5b	Isoform 1 of Sister chromatid cohesion	−1.12
				protein PDS5 homolog B

IPI:IPI00471475.1	HSGSDR{circumflex over ( )}S*SFSHYSGLK@	427	Serbp1	Isoform 1 of Plasminogen activator	−1.119
				inhibitor 1 RNA-binding protein

IPI:IPI00330240.7	S*QDAESYQNVVELK@	428	Peg3	Isoform 1 of Paternally-expressed gene	−1.108
				3 protein

IPI:IPI00153986.2	GTS*RPGTPSAEAASTSSTLR	429	Gtf2f1	General transcription factor IIF	−1.102
				subunit 1

IPI:IPI00453656.2	KVAPLS*SSLDTSLDFSK	430	Uvrag	UV radiation resistance associated	−1.098

IPI:IPI00127976.1	SVSENSLVAMDFSGQIGR	431	Grb10	Isoform 3 of Growth factor receptor-	−1.097
				bound protein 10

IPI:IPI00135443.2	KTSFDQDSDVDIFPSDFT	432	Top2b	DNA topoisomerase 2-beta	−1.092
	SEPPALPR

IPI:IPI00331612.3	KPAQETEETSSQESAEED	433	Hmga2	High mobility group protein HMGI-C	−1.078

IPI:IPI00331612.3	K@PAQETEETSSQESAE	434	Hmga2	High mobility group protein HMGI-C	−1.078
	ED

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAK	435	Eef1b2	Elongation factor 1-beta	−1.075
	@DDDDIDLFGS*DDEEESE
	EAK@

IPI:IPI00321884.1	ESLPLDLS*DDQSNSK	436	Nvl	Nuclear valosin-containing protein-like	−1.071

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	437	Npm1	Nucleophosmin	−1.071
	AES*EDEDEEDVK@LLGMS
	GK@

IPI:IPI00553798.2	GGVTGSPEASISGS*K@G	438	Ahnak	AHNAK nucleoprotein isoform 1	−1.067
	DLK@

IPI:IPI00136107.1	THSTSSSIGSGESPFSR	439	Ndrg3	Protein NDRG3	−1.054

IPI:IPI00130920.1	VLSPLR{circumflex over ( )}SPPLLGSESPYE	440	Mtap1b	Microtubule-associated protein 1B	−1.054
	DFLSADSK@

IPI:IPI00349069.4	GPQQDSDSDDGEALGGN	441	B230208H17Rik	Putative GTP-binding protein Parf	−1.048
	PMVAGFQDDVDIEDQTHGK

IPI:IPI00349069.4	GPQQDSDSDDGEALGGN	442	B230208H17Rik	Putative GTP-binding protein Parf	−1.048
	PMVAGFQDDVDIEDQTHG
	K@

IPI:IPI00408803.6	GR{circumflex over ( )}ESDEDTEDAS*ETDL	443	Suds3	Sin3 histone deacetylase corepressor	−1.047
	AK@			complex component SDS3

IPI:IPI00109311.3	SASSDTSEELNS*QDSPK	444	Slc9a3r1	Isoform 1 of Na(+)/H(+) exchange	−1.047
	@R{circumflex over ( )}			regulatory cofactor NHE-RF1

IPI:IPI00136107.1	THSTSS*SIGSGESPFSR{circumflex over ( )}	445	Ndrg3	Protein NDRG3	−1.042

IPI:IPI00114560.5	FRT*ITSSYYR	446	Rab1	Ras-related protein Rab-1A	−1.042

IPI:IPI00123410.5	TIS*AQDTLAYATALLNEK	447	Usp24	Isoform 1 of Ubiquitin carboxyl-	−1.04
				terminal hydrolase 24

IPI:IPI00169477.1	NTPSQHSHSIQHSPER{circumflex over ( )}	448	Bclaf1	Isoform 2 of Bcl-2-associated	−1.036
				transcription factor 1

IPI:IPI00317401.6	AESPETSAVESTQSTPQK	449	Pds5b	Isoform 1 of Sister chromatid cohesion	−1.036
				protein PDS5 homolog B

IPI:IPI00317401.6	METVSNASSSSNPSS*PGR{circumflex over ( )}	450	Pds5b	Isoform 1 of Sister chromatid cohesion	−1.035
				protein PDS5 homolog B

IPI:IPI00317401.6	METVSNASSSSNPSS*PGR	451	Pds5b	Isoform 1 of Sister chromatid cohesion	−1.035
				protein PDS5 homolog B

IPI:IPI00229702.1	KDSPPLVTPPQS*PPSSQ	452	Smtnl2	Smoothelin-like protein 2	−1.03
	PPAMTQAPR

IPI:IPI00107999.1	SSPS*ILAVQR	453	Chd1	Chromodomain-helicase-DNA-binding	−1.019
				protein 1

IPI:IPI00107999.1	SS*PSILAVQR{circumflex over ( )}	454	Chd1	Chromodomain-helicase-DNA-binding	−1.019
				protein 1

IPI:IPI00118120.1	TSS*IADEGTYTLDSILR	455	Myo5a	Myosin-Va	−1.018

IPI:IPI00453848.5	SAFK@LDSDSDEEDAQF	456	Atg2a	Protein	−1.014
	FSMASGVPQTPAPEPSR{circumflex over ( )}R{circumflex over ( )}

IPI:IPI00330773.3	AS*YSGTSPSHSFISGEPDR{circumflex over ( )}	457	Phldb2	Isoform 1 of Pleckstrin homology-like	−1.013
				domain family B member 2

IPI:IPI00225062.2	GCSPPKSPEKPPQSTSS*	458	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.003
	ESCPPSPQPTK			matrix protein 2

IPI:IPI00229613.3	SSS*SASQSNHSCTSMPSS	459	Macf1	Isoform 3 of Microtubule-actin cross-	−1.003
	PATPASGTK			linking factor 1

IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	460	Ndrg3	Protein NDRG3	−1.002

IPI:IPI00121135.5	R.SKSPPKSPEEEGAVS	461	Sfrs2	Splicing factor	−1
	S.

IPI:IPI00126338.5	R.SSTPLPTVSSS*AENTR	462	Tmpo	Isoform Alpha of Lamina-associated	−1
	{circumflex over ( )}.Q			polypeptide	2

IPI:IPI00623249.4	R.TDSVIIADQTPT*	463	Atf7	Activating transcription factor 7	−1
	PTR{circumflex over ( )}.F

Rapa replicate 2 (Note: heavy cells were control and light cells were

treated with rapamycin. L, light; H, heavy.)

IPI:IPI00330112.1	VAPAGSAIS*NASGER{circumflex over ( )}	464	Mib1	E3 ubiquitin-protein ligase MIB1	9.6909

IPI:IPI00108454.2	R{circumflex over ( )}LSSLR{circumflex over ( )}AS*TSK@	465	rpS6	29 kDa protein	9.6182

IPI:IPI00108454.2	R{circumflex over ( )}LSSLR{circumflex over ( )}AS*TSK@	466	rpS6	29 kDa protein	9.6182

IPI:IPI00225062.2	ELS*HSPPRENSFESSLEFK	467	Srrm2	Isoform 3 of Serine/arginine repetitive	9.4619
				matrix protein 2

IPI:IPI00108454.2	R{circumflex over ( )}LSSLR{circumflex over ( )}AS*TSK@	468	rpS6	29 kDa protein	9.4253

IPI:IPI00346155.4	NFLSMDSLK@	469	Gm13099	Novel protein similar to preferentially	9.2638
				expressed antigen in melanoma-like
				family

IPI:IPI00649156.4	NLAITGGVTLS*VIASPVI	470	Rnf19b	IBR domain containing 3	9.1171
	AAVSVGIGVPIMLAYVY
	GVVPISLCR{circumflex over ( )}

IPI:IPI00108454.2	R{circumflex over ( )}LSSLR{circumflex over ( )}AS*TSK@	471	rpS6	29 kDa protein	9.0561

IPI:IPI00133349.1	TR{circumflex over ( )}TFS*ATVR{circumflex over ( )}	472	Carhsp1	Calcium-regulated heat stable protein 1	8.7001

IPI:IPI00133349.1	TR{circumflex over ( )}TFS*ATVR{circumflex over ( )}	473	Carhsp1	Calcium-regulated heat stable protein 1	8.6779

IPI:IPI00133349.1	TR{circumflex over ( )}TFS*ATVR{circumflex over ( )}	474	Carhsp1	Calcium-regulated heat stable protein 1	8.6519

IPI:IPI00108454.2	R{circumflex over ( )}LSSLR{circumflex over ( )}AS*TSK@	475	rpS6	29 kDa protein	8.6271

IPI:IPI00351206.5	GLCSGPGAGEESPAAT*LP	476	Pgrmc2	Membrane-associated progesterone	8.5943
	R{circumflex over ( )}M#K@			receptor component	2

IPI:IPI00108454.2	LSSLR{circumflex over ( )}ASTSK@	477	rpS6	29 kDa protein	7.5465

IPI:IPI00108454.2	LSSLR{circumflex over ( )}ASTSK@	478	rpS6	29 kDa protein	7.5465

IPI:IPI00131138.1	AHVAPCFDAS*K@	479			7.3937

IPI:IPI00125530.1	WTNDYSMVLTGAAIY*HK@	480	Ext1	Exostosin-1	7.3545

IPI:IPI00108454.2	LSSLR{circumflex over ( )}ASTSK@	481	rpS6	29 kDa protein	7.1711

IPI:IPI00108454.2	LSSLR{circumflex over ( )}ASTSK@	482	rpS6	29 kDa protein	7.1711

IPI:IPI00108454.2	R{circumflex over ( )}LSSLR{circumflex over ( )}ASTSK@	483	rpS6	29 kDa protein	7.0714

IPI:IPI00131138.1	AHVAPCFDAS*K@	484			7.0413

IPI:IPI00378564.4	LVYPEVK@ESK@FK@	485	Gm14085	Novel protein similar to solute carrier	6.9768
				family 28 (Sodium-coupled nucleoside
				transporter) member 2

IPI:IPI00108454.2	R{circumflex over ( )}LSSLR{circumflex over ( )}ASTSK@	486	rpS6	29 kDa protein	6.9459

IPI:IPI00133685.1	AATATRPPGPPPAPQPPS*	487	Akt1s1	Proline-rich AKT1 substrate 1	6.9065
	PAPS*PPPRPALAR

IPI:IPI00316922.1	R{circumflex over ( )}LGAAM#T*FPR{circumflex over ( )}PPSR{circumflex over ( )}	488	Gm9757	Putative uncharacterized protein	6.6604

IPI:IPI00118875.4	ATAPQTQHVS*PMR{circumflex over ( )}	489	Eef1d	eukaryotic translation elongation	6.5442
				factor 1 delta isoform a

IPI:IPI00109318.1	R{circumflex over ( )}NSPMAQTPPCHLPNIP	490	Eif4ebp2	Eukaryotic translation initiation	6.5094
	GVTSPGALIEDSK@			factor 4E-binding protein 2

IPI:IPI00109318.1	R{circumflex over ( )}NSPMAQTPPCHLPNIP	491	Eif4ebp2	Eukaryotic translation initiation	6.4811
	GVTSPGALIEDSK@			factor 4E-binding protein 2

IPI:IPI00131138.1	AHVAPCFDAS*K@	492			6.4769

IPI:IPI00757687.1	LK@PDS*K@PFSR{circumflex over ( )}	493	Gon4l	RIKEN cDNA 5830417I10 gene	6.4593

IPI:IPI00121027.4	ESLSHVLSK@	494	Mogat1	2-acylglycerol O-acyltransferase 1	6.0819

IPI:IPI00115681.1	TECGSTGSPASSFHFK@	495	Eef2k	Elongation factor 2 kinase	5.912

IPI:IPI00115681.1	TECGSTGSPASSFHFK@	496	Eef2k	Elongation factor 2 kinase	5.912

IPI:IPI00128818.2	LYTEK@AY*K@	497	Dhx15	Putative pre-mRNA-splicing factor ATP-	5.9047
				dependent RNA helicase DHX15

IPI:IPI00118438.4	SR{circumflex over ( )}VSVSPGR{circumflex over ( )}	498	Srrm1	Isoform	2 of Serine/arginine repetitive	5.834
				matrix protein 1

IPI:IPI00323238.5	R{circumflex over ( )}K@TVSFSAAEEAPVPE	499	Setd1a	SET domain containing 1A	5.7891
	PSTAAPLQAK@

IPI:IPI00323238.5	R{circumflex over ( )}K@TVSFSAAEEAPVPE	500	Setd1a	SET domain containing 1A	5.6797
	PSTAAPLQAK@

IPI:IPI00323238.5	R{circumflex over ( )}K@TVSFSAAEEAPVPE	501	Setd1a	SET domain containing 1A	5.6797
	PSTAAPLQAK@

IPI:IPI00405470.1	SGHSLLT*WDSAMSALSSA	502	D830015G02Rik	Putative uncharacterized protein	5.6705
	S*SSSCCTLR{circumflex over ( )}

IPI:IPI00137148.2	R{circumflex over ( )}K@Y*GMLHTQESSSSVF	503	Vwa5b1	von Willebrand factor A domain-	5.6371
	YPSQDEGLSPGSGNCAK@			containing protein 5B1

IPI:IPI00660767.2	SDSSGGYTLSDVIQSPPS	504	Ibtk	Isoform 2 of Inhibitor of Bruton	5.5816
	AGLLK@			tyrosine kinase

IPI:IPI00660767.2	SDSSGGYTLSDVIQSPPS	505	Ibtk	Isoform	2 of Inhibitor of Bruton	5.4166
	AGLLK@			tyrosine kinase

IPI:IPI00351206.5	GLCSGPGAGEESPAAT*LP	506	Pgrmc2	Membrane-associated progesterone	5.4066
	R{circumflex over ( )}M#K@			receptor component 2

IPI:IPI00117731.1	TFWS*PELK@K@	507	Erc1;	Isoform 1 of ELKS/RAB6-	5.3901
			LOC100048600	interacting/CAST family member 1

IPI:IPI00115681.1	TECGSTGSPASSFHFK@	508	Eef2k	Elongation factor 2 kinase	5.3519

IPI:IPI00351206.5	GLCSGPGAGEES*PAATLP	509	Pgrmc2	Membrane-associated progesterone	5.2793
	R{circumflex over ( )}M#K@			receptor component 2

IPI:IPI00108454.2	LSSLR{circumflex over ( )}ASTSK@SESSQK@	510	rpS6	29 kDa protein	5.274

IPI:IPI00108454.2	LSSLR{circumflex over ( )}ASTSK@SESSQK@	511	rpS6	29 kDa protein	5.274

IPI:IPI00108454.2	R{circumflex over ( )}LSSLR{circumflex over ( )}	512	rpS6	29 kDa protein	5.2699

IPI:IPI00108454.2	RLSSLR	513	rpS6	29 kDa protein	5.2699

IPI:IPI00108454.2	LSSLR{circumflex over ( )}ASTSK@SESSQK@	514	rpS6	29 kDa protein	5.257

IPI:IPI00331280.1	AALDQT*SGLPGGAAQDPG	515	Adamts2	A disintegrin and metalloproteinase	5.2222
	GR{circumflex over ( )}			with thrombospondin motifs 2

IPI:IPI00108454.2	RLSSLR	516	rpS6	29 kDa protein	5.1769

IPI:IPI00108454.2	R{circumflex over ( )}LSSLR{circumflex over ( )}	517	rpS6	29 kDa protein	5.1745

IPI:IPI00127976.1	MNILSSQSPLHPSTLNAVI	518	Grb10	Isoform	3 of Growth factor receptor-	5.0363
	HR{circumflex over ( )}			bound protein 10

IPI:IPI00127976.1	MNILSSQSPLHPSTLNAVI	519	Grb10	Isoform	3 of Growth factor receptor-	5.0363
	HR{circumflex over ( )}			bound protein 10

IPI:IPI00656285.2	SLVSPIPS*PTGTISVPNSCP	520	Foxk1	Forkhead box protein K1	5.0164
	AS*PR{circumflex over ( )}

IPI:IPI00461419.2	IR{circumflex over ( )}SSLAR{circumflex over ( )}	521	Gm6988	similar to hCG1640785	4.998

IPI:IPI00108454.2	R{circumflex over ( )}LSSLR{circumflex over ( )}ASTSK@	522	rpS6	29 kDa protein	4.9653

IPI:IPI00127976.1	MNILSSQSPLHPSTLNAVI	523	Grb10	Isoform	3 of Growth factor receptor-	4.9297
	HR{circumflex over ( )}			bound protein 10

IPI:IPI00403938.2	LLDPQEFTLSGT*QR{circumflex over ( )}	524	Tnc	Isoform	1 of Tenascin	4.9074

IPI:IPI00108454.2	R{circumflex over ( )}LSSLR{circumflex over ( )}ASTSK@	525	rpS6	29 kDa protein	4.8929

IPI:IPI00929786.1	TASISSSPSEGTPAVGSY	526	Larp1	Isoform	1 of La-related protein 1	4.8902
	GCT*PQSLPK@

IPI:IPI00169986.4	SLLVLR{circumflex over ( )}GTAYTR{circumflex over ( )}	527	Alkbh6	24 kDa protein	4.8761

IPI:IPI00127976.1	MNILSSQSPLHPSTLNAVI	528	Grb10	Isoform	3 of Growth factor receptor-	4.774
	HR{circumflex over ( )}			bound protein 10

IPI:IPI00929786.1	TASISSSPSEGTPAVGSY	529	Larp1	Isoform	1 of La-related protein 1	4.758
	GCTPQSLPK@

IPI:IPI00929786.1	TASISSSPSEGTPAVGSY*	530	Larp1	Isoform	1 of La-related protein 1	4.758
	GCT*PQSLPK@

IPI:IPI00115681.1	TECGSTGSPASSFHFK@	531	Eef2k	Elongation factor 2 kinase	4.7395

IPI:IPI00929786.1	TASISSSPSEGTPAVGSY	532	Larp1	Isoform	1 of La-related protein 1	4.7282
	GCTPQSLPK@

IPI:IPI00127976.1	M#NILSSQSPLHPSTLNAV	533	Grb10	Isoform	3 of Growth factor receptor-	4.7179
	IHR{circumflex over ( )}			bound protein 10

IPI:IPI00340860.5	TASEGSEAETPEAPK@Q	534	Larp7	Isoform	1 of La-related protein 7	4.7122
	PAK@K@

IPI:IPI00340860.5	TASEGSEAETPEAPK@Q	535	Larp7	Isoform	1 of La-related protein 7	4.7105
	PAK@K@

IPI:IPI00127976.1	MNILSSQS*PLHPSTLNAVIH	536	Grb10	Isoform	3 of Growth factor receptor-	4.6623
	R{circumflex over ( )}			bound protein 10

IPI:IPI00127976.1	MNILSSQS*PLHPSTLNAVIH	537	Grb10	Isoform	3 of Growth factor receptor-	4.6623
	R{circumflex over ( )}			bound protein 10

IPI:IPI00929786.1	TASISSSPSEGTPAVGSY	538	Larp1	Isoform	1 of La-related protein 1	4.5768
	GCTPQS*LPK@

IPI:IPI00115681.1	TECGSTGSPASSFHFK@	539	Eef2k	Elongation factor 2 kinase	4.562

IPI:IPI00553798.2	VK@T*PEMIIQK@PK@	540	Ahnak	AHNAK nucleoprotein isoform 1	4.557

IPI:IPI00929786.1	TASISSSPSEGTPAVGSY*	541	Larp1	Isoform	1 of La-related protein 1	4.4981
	GCT*PQSLPK@

IPI:IPI00929786.1	TASISSSPSEGTPAVGSY	542	Larp1	Isoform	1 of La-related protein 1	4.4981
	GCTPQSLPK@

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAK	543	Eef1b2	Elongation factor 1-beta	4.4765
	@DDDDIDLFGS*DDEEESE
	EAK@K@

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAKD	544	Eef1b2	Elongation factor 1-beta	4.4765
	DDDIDLFGS*DDEEESEEAKK

IPI:IPI00474916.3	AVVPFLGVGFSSLDMS*LCV	545	Qsox2	Isoform 3 of Sulfhydryl oxidase 2	4.4747
	VLYVASSLFLM#IMYFFFR
	{circumflex over ( )}VR{circumflex over ( )}

IPI:IPI00399953.1	K@FS*APGQLCVPMTSNLG	546	Wnk1	Serine/threonine-protein kinase WNK1	4.3426
	GSTPISAASATSLGHFTK@

IPI:IPI00654415.2	MTSTDYSLSGTCGM#D	547	Megf11	Isoform 4 of Multiple epidermal growth	4.325
	R{circumflex over ( )}			factor-like domains 11

IPI:IPI00660767.2	SDSSGGYTLSDVIQSPPS	548	Ibtk	Isoform	2 of Inhibitor of Bruton	4.2681
	AGLLK@			tyrosine kinase

IPI:IPI00660767.2	K@R{circumflex over ( )}SDSSGGYT*LSDVIQS	549	Ibtk	Isoform 2 of Inhibitor of Bruton	4.2613
	*PPSAGLLK@			tyrosine kinase

IPI:IPI00127976.1	MNILSSQS*PLHPSTLNAVIH	550	Grb10	Isoform	3 of Growth factor receptor-	4.2564
	R{circumflex over ( )}			bound protein 10

IPI:IPI00127976.1	MNILSSQSPLHPS*TLNAVIH	551	Grb10	Isoform	3 of Growth factor receptor-	4.2564
	R{circumflex over ( )}			bound protein 10

IPI:IPI00127976.1	M#NILSSQS*PLHPSTLNAVI	552	Grb10	Isoform	3 of Growth factor receptor-	4.2332
	HR{circumflex over ( )}			bound protein 10

IPI:IPI00313653.2	QFCQEGQPHVLEALS*PPQ	553	Maf1	Repressor of RNA polymerase III	4.2077
	TSGLS*PSR{circumflex over ( )}			transcription MAF1 homolog

IPI:IPI00929786.1	TASISSSPSEGTPAVGSY	554	Larp1	Isoform	1 of La-related protein 1	4.18
	GCTPQSLPK@

IPI:IPI00127976.1	M#NILSSQS*PLHPSTLNAVI	555	Grb10	Isoform	3 of Growth factor receptor-	4.1504
	HR{circumflex over ( )}			bound protein 10

IPI:IPI00127976.1	M#NILSSQS*PLHPSTLNAVI	556	Grb10	Isoform	3 of Growth factor receptor-	4.1504
	HR{circumflex over ( )}			bound protein 10

IPI:IPI00122594.4	EVSVSSVTEEPK@LDSSQ	557	Ahctf1	AT-hook-containing transcription	4.144
	LPLQTGLDVPAT*PR{circumflex over ( )}			factor 1

IPI:IPI00127976.1	MNILSSQS*PLHPSTLNAVIH	558	Grb10	Isoform	3 of Growth factor receptor-	4.133
	R{circumflex over ( )}			bound protein 10

IPI:IPI00127976.1	MNILSSQS*PLHPSTLNAVIH	559	Grb10	Isoform	3 of Growth factor receptor-	4.133
	R{circumflex over ( )}			bound protein 10

IPI:IPI00109318.1	R{circumflex over ( )}NSPMAQTPPCHLPNIP	560	Eif4ebp2	Eukaryotic translation initiation	4.0914
	GVTSPGALIEDSK@			factor 4E-binding protein 2

IPI:IPI00109318.1	R{circumflex over ( )}NSPMAQTPPCHLPNIP	561	Eif4ebp2	Eukaryotic translation initiation	4.0914
	GVTSPGALIEDSK@			factor 4E-binding protein 2

IPI:IPI00122521.1	K@DELSDWS*LAGEDDR{circumflex over ( )}E	562	Fxr1	Isoform E of Fragile X mental	4.0773
	TR{circumflex over ( )}			retardation syndrome-related protein 1

IPI:IPI00127976.1	MNILSS*QSPLHPSTLNAV	563	Grb10	Isoform	3 of Growth factor receptor-	4.0585
	IHR			bound protein 10

IPI:IPI00127976.1	MNILSSQSPLHPSTLNAVI	564	Grb10	Isoform	3 of Growth factor receptor-	4.0417
	HR{circumflex over ( )}			bound protein 10

IPI:IPI00313653.2	QFCQEGQPHVLEALS*PPQ	565	Maf1	Repressor of RNA polymerase III	4.0213
	TSGLS*PSR{circumflex over ( )}			transcription MAF1 homolog

IPI:IPI00127976.1	MNILSS*QSPLHPSTLNAV	566	Grb10	Isoform	3 of Growth factor receptor-	3.9734
	IHR			bound protein 10

IPI:IPI00127976.1	MNILSSQS*PLHPSTLNAVIH	567	Grb10	Isoform	3 of Growth factor receptor-	3.9734
	R{circumflex over ( )}			bound protein 10

IPI:IPI00127976.1	MNILSSQS*PLHPSTLNAVIH	568	Grb10	Isoform	3 of Growth factor receptor-	3.9734
	R{circumflex over ( )}			bound protein 10

IPI:IPI00127976.1	MNILSS*QSPLHPSTLNAV	569	Grb10	Isoform	3 of Growth factor receptor-	3.9734
	IHR			bound protein 10

IPI:IPI00120691.3	QADSETK@EIITEEPSEEE	570	Ddx21	Nucleolar RNA helicase 2	3.9589
	ADMPK@PK@

IPI:IPI00226441.2	LHYTPPLQSPIT*DGDPLL	571	Lin9	Isoform 2 of Lin-9 homolog	3.8536
	GQS*PWR{circumflex over ( )}

IPI:IPI00127976.1	MNILSSQS*PLHPSTLNAVIH	572	Grb10	Isoform	3 of Growth factor receptor-	3.8383
	R{circumflex over ( )}			bound protein 10

IPI:IPI00127976.1	MNILSSQS*PLHPSTLNAVIH	573	Grb10	Isoform	3 of Growth factor receptor-	3.8383
	R{circumflex over ( )}			bound protein 10

IPI:IPI00656285.2	SSGLQTPECLSREGSPIP	574	Foxk1	Forkhead box protein K1	3.7948
	HDPDLGSK

IPI:IPI00656285.2	SSGLQTPECLSR{circumflex over ( )}EGSPIP	575	Foxk1	Forkhead box protein K1	3.7948
	HDPDLGSK@

IPI:IPI00468516.3	SHILESVPTLPGS*VEAGVS	576	D6Wsu116e	Isoform 1 of Protein FAM21	3.7932
	FDLPAQADTLHSANK@

IPI:IPI00751009.1	TSEFPTPLFSGPLEPVACGS	577	Srpk2	serine/arginine-rich protein-specific	3.7653
	VISEGSPLTEQEESSPSHDR			kinase 2
	{circumflex over ( )}S*R{circumflex over ( )}

IPI:IPI00929786.1	TASISSSPS*EGTPAVGSYG	578	Larp1	Isoform	1 of La-related protein 1	3.756
	CTPQSLPK@

IPI:IPI00929786.1	TASISSSPSEGTPAVGSY	579	Larp1	Isoform	1 of La-related protein 1	3.756
	GCT*PQSLPK@

IPI:IPI00656285.2	SSGLQTPECLSR{circumflex over ( )}EGSPIP	580	Foxk1	Forkhead box protein K1	3.7214
	HDPDLGSK@

IPI:IPI00400017.2	TQS*GNFNTDAPGMAEFR{circumflex over ( )}	581	Ppfibp2	Isoform	4 of Liprin-beta-2	3.6603

IPI:IPI00660767.2	SDSSGGYTLSDVIQSPPS	582	Ibtk	Isoform 2 of Inhibitor of Bruton	3.6519
	AGLLK@			tyrosine kinase

IPI:IPI00116442.1	S*EGLSLER{circumflex over ( )}	583	Hdgfrp2	Isoform	3 of Hepatoma-derived growth	3.6501
				factor-related protein 2

IPI:IPI00379844.4	TAS*EGDGGAAGGAGTAG	584	Irs2	Insulin receptor substrate 2	3.6492
	GR{circumflex over ( )}PMSVAGSPLSPGPVR{circumflex over ( )}

IPI:IPI00929786.1	TASISSSPSEGTPAVGS*Y	585	Larp1	Isoform	1 of La-related protein 1	3.6217
	GCTPQSLPK@

IPI:IPI00929786.1	TASISSSPSEGTPAVGS*Y	586	Larp1	Isoform	1 of La-related protein 1	3.6217
	GCTPQSLPK@

IPI:IPI00116442.1	R{circumflex over ( )}S*EGLSLER{circumflex over ( )}	587	Hdgfrp2	Isoform	3 of Hepatoma-derived growth	3.6142
				factor-related protein 2

IPI:IPI00656285.2	SLVSPIPSPTGTISVPNSC	588	Foxk1	Forkhead box protein K1	3.6065
	PAS*PR{circumflex over ( )}

IPI:IPI00656285.2	SLVSPIPSPTGTISVPNSC	589	Foxk1	Forkhead box protein K1	3.6065
	PAS*PR{circumflex over ( )}

IPI:IPI00656285.2	SLVSPIPSPT*GTISVPNSCP	590	Foxk1	Forkhead box protein K1	3.605
	AS*PR{circumflex over ( )}

IPI:IPI00654417.1	HSS*PPLTVK	591	Tinf2	Putative uncharacterized protein	3.5562

IPI:IPI00830829.1	R{circumflex over ( )}R{circumflex over ( )}SGEQITSSPVS*PK@	592	Hisppd1	140 kDa protein	3.5492

IPI:IPI00118875.4	GATPAEDDEDKDIDLFGS*D	593	Eef1d	eukaryotic translation elongation	3.5469
	EEEEDKEAAR			factor 1 delta isoform a

IPI:IPI00118875.4	GATPAEDDEDKDIDLFGS*D	594	Eef1d	eukaryotic translation elongation	3.5469
	EEEEDKEAAR			factor 1 delta isoform a

IPI:IPI00169888.3	SQDVAISPQQQQCSK@S	595	Edc3	Enhancer of mRNA-decapping protein 3	3.5277
	YVDR{circumflex over ( )}

IPI:IPI00134029.8	VSSSVSVGPSM#SGETL	596	Aim1l	Absent in melanoma 1-like	3.5092
	PR{circumflex over ( )}

IPI:IPI00654417.1	HSS*PPLTVK	597	Tinf2	Putative uncharacterized protein	3.4997

IPI:IPI00676574.2	RVS*TDLPEGQDVYTAACN	598	Herc1	hect (homologous to the E6-AP	3.4866
	SVIHR			(UBE3A) carboxyl terminus) domain
				and RCC1 (CHC1)-like domain (RLD) 1

IPI:IPI00762234.2	SGDPR{circumflex over ( )}SCEEESDPEPDP	599	Fbxw9	F-box and WD-40 domainprotein 9	3.4479
	DPDTQAEAYVAR{circumflex over ( )}

IPI:IPI00660767.2	VDTAASSSWLAGSCSPVS	600	Ibtk	Isoform	2 of Inhibitor of Bruton	3.398
	PPVVDLR{circumflex over ( )}			tyrosine kinase

IPI:IPI00169888.3	SQDVAISPQQQQCSK@S	601	Edc3	Enhancer of mRNA-decapping protein 3	3.3665
	YVDR{circumflex over ( )}

IPI:IPI00169888.3	SQDVAISPQQQQCS*K@SY	602	Edc3	Enhancer of mRNA-decapping protein 3	3.3665
	*VDR{circumflex over ( )}

IPI:IPI00122521.1	K@DELSDWS*LAGEDDR{circumflex over ( )}E	603	Fxr1	Isoform E of Fragile X mental	3.3379
	TR{circumflex over ( )}			retardation syndrome-related protein 1

IPI:IPI00761443.2	RSSAGSLESNVEGSIISS*	604	Iqsec1	IQ motif and Sec7 domain 1 isoform b	3.3179
	PHMR

IPI:IPI00761443.2	R{circumflex over ( )}SSAGSLESNVEGSIISS*	605	Iqsec1	IQ motif and Sec7 domain 1 isoform b	3.3179
	PHMR{circumflex over ( )}

IPI:IPI00828461.1	GPPDFSSDEER{circumflex over ( )}EPTPVL	606	Tmpo	thymopoietin isoform delta	3.3038
	GSGASVGR{circumflex over ( )}

IPI:IPI00828461.1	GPPDFSSDEER{circumflex over ( )}EPTPVL	607	Tmpo	thymopoietin isoform delta	3.3038
	GSGASVGR{circumflex over ( )}

IPI:IPI00126317.1	NASTSFQELEDK@K@EL	608	Dnajc2	DnaJ homolog subfamily C member 2	3.2768
	SEESEDEELQLEEFPMLK@

IPI:IPI00227934.3	TSASCSPAPESPMSSSES	609	Cux1	cut-like homeobox 1 isoform a	3.1887
	VK@

IPI:IPI00116442.1	S*EGLSLER{circumflex over ( )}	610	Hdgfrp2	Isoform	3 of Hepatoma-derived growth	3.1751
				factor-related protein 2

IPI:IPI00468516.3	SHILESVPTLPGS*VEAGVS	611	D6Wsu116e	Isoform	1 of Protein FAM21	3.1635
	FDLPAQADTLHSANK@

IPI:IPI00468516.3	SHILESVPTLPGS*VEAGVS	612	D6Wsu116e	Isoform	1 of Protein FAM21	3.1635
	FDLPAQADTLHSANK@

IPI:IPI00762234.2	SGDPR{circumflex over ( )}SCEEESDPEPDP	613	Fbxw9	F-box and WD-40 domain protein 9	3.1508
	DPDTQAEAYVAR{circumflex over ( )}

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAKD	614	Eef1b2	Elongation factor 1-beta	3.1436
	DDDIDLFGS*DDEEESEEAKK

IPI:IPI00320594.5	SQPHSSTSNQETSDSEM	615	Ranbp10	Ran-binding protein 10	3.1377
	EMEAEHYPNGVLESVSTR{circumflex over ( )}

IPI:IPI00808277.2	S*APASPNHAGVLSAHSSG	616	Foxk2	Isoform	1 of Forkhead box protein K2	3.1204
	AQTPESLSR{circumflex over ( )}EGSPAPLE
	PEPGASQPK@

IPI:IPI00808277.2	SAPAS*PNHAGVLSAHSSG	617	Foxk2	Isoform 1 of Forkhead box protein K2	3.1105
	AQTPESLSR{circumflex over ( )}EGSPAPLE
	PEPGASQPK@

IPI:IPI00753321.2	SVSETSEDK@K@DEESD	618	Bod1l	biorientation of chromosomes in cell	3.106
	EEEEEEEEEEPLGATTR{circumflex over ( )}			division 1-like

IPI:IPI00420464.3	SNTPSPLNLSSTSSK@	619	Zeb2	Zinc finger E-box-binding homeobox 2	3.0996

IPI:IPI00116442.1	S*EGLSLER{circumflex over ( )}	620	Hdgfrp2	Isoform	3 of Hepatoma-derived growth	3.0974
				factor-related protein 2

IPI:IPI00808277.2	SAPAS*PNHAGVLSAHSSG	621	Foxk2	Isoform	1 of Forkhead box protein K2	3.0877
	AQTPESLSR{circumflex over ( )}

IPI:IPI00830829.1	R{circumflex over ( )}R{circumflex over ( )}SGEQITSSPVS*PK@	622	Hisppd1	140 kDa protein	3.0712

IPI:IPI00117932.2	S*PPVQPHTPVTISLGTAPS	623	Sin3a	Isoform	1 of Paired amphipathic helix	3.0565
	LQNNQPVEFNHAINYVNK@			protein Sin3a

IPI:IPI00117932.2	S*PPVQPHTPVTISLGTAPS	624	Sin3a	Isoform	1 of Paired amphipathic helix	3.0565
	LQNNQPVEFNHAINYVNK@			protein Sin3a

IPI:IPI00320594.5	SQPHSSTSNQETSDSEM	625	Ranbp10	Ran-binding protein 10	3.0328
	EMEAEHYPNGVLESVSTR{circumflex over ( )}

IPI:IPI00227934.3	TSASCSPAPESPMSSSES	626	Cux1	cut-like homeobox 1 isoform a	3.0118
	VK@

IPI:IPI00380817.6	GR{circumflex over ( )}SS*ESSCGLDGDYEDA	627	Bcr	Breakpoint cluster region protein	3.0087
	ELNPR{circumflex over ( )}

IPI:IPI00656285.2	SLVSPIPS*PTGTISVPNSCP	628	Foxk1	Forkhead box protein K1	2.9994
	AS*PR{circumflex over ( )}

IPI:IPI00656285.2	SLVSPIPSPTGTISVPNSC	629	Foxk1	Forkhead box protein K1	2.9989
	PAS*PR{circumflex over ( )}

IPI:IPI00127976.1	MNILSSQSPLHPSTLNAVI	630	Grb10	Isoform	3 of Growth factor receptor-	2.9784
	HR{circumflex over ( )}			bound protein 10

IPI:IPI00551454.3	ER{circumflex over ( )}QES*ESEQELVNK@	631	Pdcd11	Protein RRP5 homolog	2.9603

IPI:IPI00226155.3	VSALEEDMDDVESSEEEE	632	Prpf38a	Isoform	1 of Pre-mRNA-splicing factor	2.9547
	EEDEKLER			38A

IPI:IPI00753321.2	SVSETSEDK@K@DEESD	633	Bod1l	biorientation of chromosomes in cell	2.9525
	EEEEEEEEEEPLGATTR{circumflex over ( )}			division 1-like

IPI:IPI00753321.2	SVSETSEDK@K@DEESD	634	Bod1l	biorientation of chromosomes in cell	2.9525
	EEEEEEEEEEPLGATTR{circumflex over ( )}			division 1-like

IPI:IPI00108454.2	LSSLR{circumflex over ( )}ASTSK@S*ESSQ	635	—	29 kDa protein	2.9394
	K@

IPI:IPI00553798.2	HR{circumflex over ( )}SNSFSDER{circumflex over ( )}EFSAPS*	636	Ahnak	AHNAK nucleoprotein isoform 1	2.9368
	TPTGTLEFAGGDAK@

IPI:IPI00551454.3	ER{circumflex over ( )}QES*ESEQELVNK@	637	Pdcd11	Protein RRP5 homolog	2.9308

IPI:IPI00468516.3	SHILESVPTLPGS*VEAGVS	638	D6Wsu116e	Isoform 1 of Protein FAM21	2.9286
	FDLPAQADTLHSANK@

IPI:IPI00420464.3	SNTPSPLNLSSTSSK@	639	Zeb2	Zinc finger E-box-binding homeobox 2	2.9204

IPI:IPI00656285.2	SLVSPIPSPTGTISVPNSC	640	Foxk1	Forkhead box protein K1	2.8837
	PAS*PR{circumflex over ( )}

IPI:IPI00330773.3	K@MLLASTSSDDFDR{circumflex over ( )}AS*	641	Phldb2	Isoform	1 of Pleckstrin homology-like	2.8699
	Y*SGTSPSHSFISGEPDR{circumflex over ( )}			domain family B member 2

IPI:IPI00551454.3	ER{circumflex over ( )}QES*ESEQELVNK@	642	Pdcd11	Protein RRP5 homolog	2.8488

IPI:IPI00656285.2	SLVSPIPSPTGTISVPN	643	Foxk1	Forkhead box protein K1	2.818
	S*CPASPR{circumflex over ( )}

IPI:IPI00660767.2	VDTAASSSWLAGSCSPVS	644	Ibtk	Isoform 2 of Inhibitor of Bruton	2.8109
	PPVVDLR{circumflex over ( )}			tyrosine kinase

IPI:IPI00656285.2	SLVSPIPSPTGTISVPNSC	645	Foxk1	Forkhead box protein K1	2.8019
	PAS*PR{circumflex over ( )}

IPI:IPI00656285.2	SLVSPIPSPTGTISVPNSC	646	Foxk1	Forkhead box protein K1	2.8019
	PAS*PR{circumflex over ( )}

IPI:IPI00656285.2	SLVSPIPSPTGTISVPNSC	647	Foxk1	Forkhead box protein K1	2.8019
	PAS*PR

IPI:IPI00107958.1	TSDIFGS*PVTATAPLAHPN	648	Hn1l	Hematological and neurological	2.7751
	K@PK@			expressed 1-like protein

IPI:IPI00129298.1	EIDVLEFGESAPAASKENSA	649	Palm	Isoform	1 of Paralemmin	2.7684
	APS*PGRPQSASPAKEEQK

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAK	650	Eef1b2	Elongation factor 1-beta	2.7577
	@DDDDIDLFGS*DDEEESE
	EAK@K@

IPI:IPI00929786.1	TASISSSPSEGTPAVGSY	651	Larp1	Isoform	1 of La-related protein 1	2.7577
	GCTPQSLPK@

IPI:IPI00377615.2	R{circumflex over ( )}SSR{circumflex over ( )}SFSLDEPPLFIPD	652	Phf3	PHD finger protein 3	2.7452
	NIATVK@

IPI:IPI00113563.3	YMEDSTYYK@ASK@	653	Ptk2	Isoform	1 of Focal adhesion kinase 1	2.7394

IPI:IPI00553283.1	MMPS*PSDSSR{circumflex over ( )}	654	1110013L07Rik	Putative uncharacterized protein	2.7037

IPI:IPI00314502.5	VHGLPTTSPS*GVNMAELA	655	Tcfeb	Transcription factor EB	2.6958
	QQVVK@

IPI:IPI00314502.5	VHGLPTTSPS*GVNMAELA	656	Tcfeb	Transcription factor EB	2.6958
	QQVVK@

IPI:IPI00122594.4	EVSVSSVTEEPK@LDSSQ	657	Ahctf1	AT-hook-containing transcription	2.6674
	LPLQTGLDVPAT*PR{circumflex over ( )}			factor 1

IPI:IPI00377615.2	SSR{circumflex over ( )}SFSLDEPPLFIPDNIA	658	Phf3	PHD finger protein 3	2.6537
	TVK@K@

IPI:IPI00468516.3	SHILESVPTLPGS*VEAGVS	659	D6Wsu116e	Isoform	1 of Protein FAM21	2.6436
	FDLPAQADTLHSANK@

IPI:IPI00122521.1	SSISSVLK@DPDSNPYSLLD	660	Fxr1	Isoform E of Fragile X mental	2.618
	NTESDQTADTDAS*ESHH			retardation syndrome-related protein 1
	STNR{circumflex over ( )}

IPI:IPI00625723.2	TGTGSPFAGNS*PAR{circumflex over ( )}EGE	661	Zc3h4	Isoform	2 of Zinc finger CCCH domain-	2.5583
	QDAGSLK@DVFK@			containing protein	4

IPI:IPI00625723.2	TGTGSPFAGNS*PAR{circumflex over ( )}EGE	662	Zc3h4	Isoform	2 of Zinc finger CCCH domain-	2.5583
	QDAGSLK@DVFK@			containing protein	4

IPI:IPI00122559.3	EIQNGTLR{circumflex over ( )}ES*DSEHVPR{circumflex over ( )}	663	Ktn1	isoform	1 of Kinectin	2.5547

IPI:IPI00136379.1	GGGSAAAAAAAAASGGGV	664	Pbx2	Pre-B-cell leukemia transcription	2.554
	S*PDNSIEHSDYR{circumflex over ( )}		factor 2

IPI:IPI00330246.2	SGPLPSSSGSSSSSSQLSV	665	Phldb1	Isoform	2 of Pleckstrin homology-like	2.5309
	ATLGR{circumflex over ( )}SPSPK@			domain family B member 1

IPI:IPI00330246.2	SGPLPSSSGSSSSSSQLSV	666	Phldb1	Isoform	2 of Pleckstrin homology-like	2.5309
	ATLGR{circumflex over ( )}SPSPK@			domain family B member 1

IPI:IPI00377615.2	SSR{circumflex over ( )}SFSLDEPPLFIPDNIA	667	Phf3	PHD finger protein 3	2.5121
	TVK@K@

IPI:IPI00125505.2	TTGNSHWTLEAPFSSS*CA	668	Cdgap	Cdc42 GTPase-activating protein	2.4733
	NLETER{circumflex over ( )}

IPI:IPI00551454.3	ER{circumflex over ( )}QES*ESEQELVNK@	669	Pdcd11	Protein RRP5 homolog	2.4392

IPI:IPI00320594.5	SQPHSSTSNQETSDSEM	670	Ranbp10	Ran-binding protein 10	2.4384
	EMEAEHYPNGVLESVSTR{circumflex over ( )}

IPI:IPI00126317.1	NASTSFQELEDK@K@EL	671	Dnajc2	DnaJ homolog subfamily C member 2	2.438
	SEESEDEELQLEEFPMLK@

IPI:IPI00129276.2	LES*LNIQR{circumflex over ( )}	672	Eif3a	Eukaryotic translation initiation	2.429
				factor 3 subunit A

IPI:IPI00553798.2	SSEVVLSGDDEDYQR{circumflex over ( )}	673	Ahnak	AHNAK nucleoprotein isoform 1	2.4187

IPI:IPI00226750.2	TSS*TCSNESLNAGGTPV	674	Tbc1d4	140 kDa protein	2.4185
	TPR{circumflex over ( )}

IPI:IPI00377615.2	R{circumflex over ( )}SSR{circumflex over ( )}SFSLDEPPLFIPD	675	Phf3	PHD finger protein 3	2.409
	NIATVK@

IPI:IPI00377615.2	R{circumflex over ( )}SSR{circumflex over ( )}SFSLDEPPLFIPD	676	Phf3	PHD finger protein 3	2.409
	NIATVK@

IPI:IPI00114227.7	SIST*CGPLDK@DDPGGQK@	677	Dennd4a	hypothetical protein LOC102442	2.3733

IPI:IPI00339693.1	LGDSELALVCSQR{circumflex over ( )}PAS*LS	678	Wdr91	WD repeat-containing protein 91	2.3532
	QSPR{circumflex over ( )}

IPI:IPI00137166.1	SR{circumflex over ( )}DATPPVSPINMEDQE	679	Junb	Transcription factor jun-B	2.3444
	R{circumflex over ( )}

IPI:IPI00751009.1	TVSASSTGDLPK@	680	Srpk2	serine/arginine-rich protein-specific	2.3444
				kinase 2

IPI:IPI00751009.1	TVSASSTGDLPK@	681	Srpk2	serine/arginine-rich protein-specific	2.3444
				kinase 2

IPI:IPI00751009.1	TVSASSTGDLPK	682	Srpk2	serine/arginine-rich protein-specific	2.3444
				kinase 2

IPI:IPI00226750.2	TSSTCSNESLNAGGTPVT	683	Tbc1d4	140 kDa protein	2.3429
	PR{circumflex over ( )}

IPI:IPI00221581.1	SR{circumflex over ( )}TGSESSQTGASATSG	684	Eif4b	Eukaryotic translation initiation	2.3268
	R{circumflex over ( )}			factor 4B

IPI:IPI00122521.1	SSISSVLK@DPDSNPYSLLD	685	Fxr1	Isoform E of Fragile X mental	2.3266
	NTESDQTADT*DASESHH			retardation syndrome-related protein 1
	STNR{circumflex over ( )}

IPI:IPI00116442.1	K@R{circumflex over ( )}SEGLSLER{circumflex over ( )}	686	Hdgfrp2	Isoform	3 of Hepatoma-derived growth	2.3042
				factor-related protein 2

IPI:IPI00828461.1	GPPDFSSDEER{circumflex over ( )}EPT*PVL	687	Tmpo	thymopoietin isoform delta	2.3011
	GS*GASVGR{circumflex over ( )}

IPI:IPI00463493.1	S*ATDADMVNSGWLVVGK	688	Gbf1	Golgi-specific brefeldin A-resistance	2.294
				factor 1

IPI:IPI00129276.2	LES*LNIQR{circumflex over ( )}	689	Eif3a	Eukaryotic translation initiation	2.2772
				factor 3 subunit A

IPI:IPI00751009.1	TVSASSTGDLPK@	690	Srpk2	serine/arginine-rich protein-specific	2.2686
				kinase 2

IPI:IPI00137166.1	SR{circumflex over ( )}DATPPVSPINMEDQE	691	Junb	Transcription factor jun-B	2.2666
	R{circumflex over ( )}

IPI:IPI00221581.1	SR{circumflex over ( )}TGSESSQTGASATSG	692	Eif4b	Eukaryotic translation initiation	2.2646
	R{circumflex over ( )}			factor 4B

IPI:IPI00221581.1	SR{circumflex over ( )}TGSESSQTGASATSG	693	Eif4b	Eukaryotic translation initiation	2.2646
	R{circumflex over ( )}			factor 4B

IPI:IPI00221581.1	SRTGSESSQTGASATSGR	694	Eif4b	Eukaryotic translation initiation	2.2646
				factor 4B

IPI:IPI00268688.3	TSSSETEEK@K@TEK@P	695	Cd2ap	CD2-associated protein	2.2567
	LILQPLGSR{circumflex over ( )}

IPI:IPI00268688.3	TSSSETEEKKTEKPLILQP	696	Cd2ap	CD2-associated protein	2.2567
	LGSR

IPI:IPI00226750.2	TSSTCSNES*LNAGGTPV	697	Tbc1d4	140 kDa protein	2.2548
	TPR{circumflex over ( )}

IPI:IPI00221581.1	SRTGSESSQTGASATSGR	698	Eif4b	Eukaryotic translation initiation	2.2395
				factor 4B

IPI:IPI00221581.1	SR{circumflex over ( )}TGSESSQTGASATSG	699	Eif4b	Eukaryotic translation initiation	2.2395
	R{circumflex over ( )}			factor 4B

IPI:IPI00122594.4	EVSVSSVTEEPK@LDSSQ	700	Ahctf1	AT-hook-containing transcription	2.2291
	LPLQTGLDVPATPR{circumflex over ( )}			factor 1

IPI:IPI00122594.4	EVSVSSVTEEPKLDSSQL	701	Ahctf1	AT-hook-containing transcription	2.2291
	PLQTGLDVPATPR			factor	1

IPI:IPI00676574.2	RVST*DLPEGQDVYTAACN	702	Herc1	hect (homologous to the E6-AP	2.2125
	SVIHR			(UBE3A) carboxyl terminus) domain
				and RCC1 (CHC1)-like domain (RLD) 1

IPI:IPI00268688.3	TSSSETEEK@K@TEK@P	703	Cd2ap	CD2-associated protein	2.1992
	LILQPLGSR{circumflex over ( )}

IPI:IPI00320905.7	LGEQGPEPGPTPPQTPTP	704	Arhgap17	Isoform 1 of Rho GTPase-activating	2.1798
	PS*TPPLAK			protein 17

IPI:IPI00108454.2	LSSLRASTSKSESSQK	705	—	29 kDa protein	2.1655

IPI:IPI00929786.1	TASISSSPSEGTPAVGSY	706	Larp1	Isoform	1 of La-related protein 1	2.1557
	GCT*PQSLPK@

IPI:IPI00381495.5	TVSSPIPYTPSPSSSR{circumflex over ( )}PIS*	707	Ccdc6	coiled-coil domain containing 6	2.1555
	PGLSY*ASHTVGFTPPTSLT
	R{circumflex over ( )}

IPI:IPI00221581.1	TGSESSQTGASATSGR{circumflex over ( )}	708	Eif4b	Eukaryotic translation initiation	2.1437
				factor 4B

IPI:IPI00221581.1	TGSESSQTGASATSGR{circumflex over ( )}	709	Eif4b	Eukaryotic translation initiation	2.1437
				factor 4B

IPI:IPI00137166.1	SR{circumflex over ( )}DATPPVSPINMEDQE	710	Junb	Transcription factor jun-B	2.1419
	R{circumflex over ( )}

IPI:IPI00929786.1	SLPTTVPESPNYR{circumflex over ( )}	711	Larp1	Isoform	1 of La-related protein 1	2.1397

IPI:IPI00929786.1	SLPTTVPESPNYR	712	Larp1	Isoform 1 of La-related protein 1	2.1397

IPI:IPI00406045.3	SESSAGICVPLSTS*PQVSE	713	Pik3r4	Phosphoinositide 3-kinase regulatory	2.1394
	AAHIPSK@K@PVIPVVSST			subunit 4
	VLPSTYQIR{circumflex over ( )}

IPI:IPI00122594.4	ER{circumflex over ( )}EVSVSSVTEEPK@LD	714	Ahctf1	AT-hook-containing transcription	2.1312
	SSQLPLQTGLDVPATPR{circumflex over ( )}			factor 1

IPI:IPI00929786.1	SLPTTVPESPNYR{circumflex over ( )}	715	Larp1	Isoform	1 of La-related protein 1	2.1207

IPI:IPI00929786.1	SLPTTVPESPNYR	716	Larp1	Isoform	1 of La-related protein 1	2.1207

IPI:IPI00929786.1	SLPTTVPESPNYR{circumflex over ( )}	717	Larp1	Isoform	1 of La-related protein 1	2.1207

IPI:IPI00929786.1	SLPTTVPESPNYR	718	Larp1	Isoform	1 of La-related protein 1	2.1207

IPI:IPI00625723.2	TGTGSPFAGNS*PAR{circumflex over ( )}EGE	719	Zc3h4	Isoform	2 of Zinc finger CCCH domain-	2.1078
	QDAGSLK@DVFK@			containing protein	4

IPI:IPI00221581.1	TGSESSQTGASATSGR{circumflex over ( )}	720	Eif4b	Eukaryotic translation initiation	2.103
				factor 4B

IPI:IPI00221581.1	TGSESSQTGASATSGR{circumflex over ( )}	721	Eif4b	Eukaryotic translation initiation	2.103
				factor 4B

IPI:IPI00221581.1	TGSESSQTGASATSGR	722	Eif4b	Eukaryotic translation initiation	2.103
				factor 4B

IPI:IPI00625723.2	TGTGSPFAGNS*PAR{circumflex over ( )}EGE	723	Zc3h4	Isoform	2 of Zinc finger CCCH domain-	2.0988
	QDAGSLK@DVFK@			containing protein	4

IPI:IPI00379844.4	GEQGSLAQSQPQPGDK@N	724	Irs2	Insulin receptor substrate 2	2.0959
	S*WSR{circumflex over ( )}

IPI:IPI00136107.1	THSTSSS*IGSGESPFSR{circumflex over ( )}	725	Ndrg3	Protein NDRG3	2.0759

IPI:IPI00467843.2	GGGASSPAPVVFTVGSPP	726	Ulk1	Putative uncharacterized protein	2.0752
	S*GATPPQSTR{circumflex over ( )}

IPI:IPI00127415.1	LK@CGSGPVHISGQHLVAV	727	Npm1	Nucleophosmin	2.0591
	EEDAES*EDEDEEDVK@LL
	GM#SGK@

IPI:IPI00320905.7	LGEQGPEPGPTPPQTPTP	728	Arhgap17	Isoform 1 of Rho GTPase-activating	2.0572
	PS*TPPLAK			protein 17

IPI:IPI00622946.2	QFIAAQNLGPASGLPTPTSS	729	Palld	Isoform 4 of Palladin	2.0517
	PSSSSLPSPLSPTPR{circumflex over ( )}PFG
	R{circumflex over ( )}

IPI:IPI00467843.2	GGGASSPAPVVFTVGSPP	730	Ulk1	Putative uncharacterized protein	2.0359
	S*GATPPOSTR{circumflex over ( )}

IPI:IPI00929786.1	TASISSSPS*EGTPAVGSYG	731	Larp1	Isoform	1 of La-related protein 1	2.0315
	CT*PQSLPK@

IPI:IPI00309059.7	STSPIIGS*PPVR{circumflex over ( )}	732	Patl1	Protein PAT1 homolog 1	2.0311

IPI:IPI00929786.1	TASISSSPSEGTPAVGSY	733	Larp1	Isoform	1 of La-related protein 1	2.0277
	GCTPQSLPK@

IPI:IPI00107958.1	TSDIFGS*PVTATAPLAHPN	734	Hn1l	Hematological and neurological	2.0059
	KPK			expressed 1-like protein

IPI:IPI00336713.1	CSPVPGLSSSPSGSPLHG	735	Bcas3	Isoform	1 of Breast carcinoma-	1.9848
	K@			amplified sequence 3 homolog

IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	736	Ndrg3	Protein NDRG3	1.9707

IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	737	Ndrg3	Protein NDRG3	1.9707

IPI:IPI00830210.1	LSESPAS*LPSCLPVETALIN	738	Atg2a	Autophagy-related protein 2	1.9651
	QR{circumflex over ( )}			homolog A

IPI:IPI00225062.2	GCSPPK@S*PEK@PPQST	739	Srrm2	Isoform	3 of Serine/arginine repetitive	1.9354
	SSESCPPSPQPTK@			matrix protein	2

IPI:IPI00122594.4	ER{circumflex over ( )}EVSVSSVTEEPK@LD	740	Ahctf1	AT-hook-containing transcription	1.9159
	SSQLPLQTGLDVPATPR{circumflex over ( )}			factor 1

IPI:IPI00336973.2	GLNLDGTPALSTLGGFS*PA	741	Ccnl1	Isoform	1 of Cyclin-L1	1.8986
	SKPSS*PR

IPI:IPI00336973.2	GLNLDGTPALSTLGGFS*PA	742	Ccnl1	Isoform 1 of Cyclin-L1	1.896
	SKPSS*PR

IPI:IPI00336973.2	GLNLDGTPALSTLGGFS*PA	743	Ccnl1	Isoform	1 of Cyclin-L1	1.896
	SK@PS*SPR{circumflex over ( )}

IPI:IPI00336973.2	GLNLDGTPALSTLGGFS*PA	744	Ccnl1	Isoform	1 of Cyclin-L1	1.896
	S*KPSSPR

IPI:IPI00336973.2	GLNLDGTPALSTLGGFSPA	745	Ccnl1	Isoform	1 of Cyclin-L1	1.896
	SK@PSSPR{circumflex over ( )}

IPI:IPI00137166.1	SR{circumflex over ( )}DATPPVSPINMEDQE	746	Junb	Transcription factor jun-B	1.8916
	R{circumflex over ( )}

IPI:IPI00336973.2	GLNLDGTPALSTLGGFS*PA	747	Ccnl1	Isoform	1 of Cyclin-L1	1.8893
	S*KPSSPR

IPI:IPI00467930.2	LVIGS*LPAHLSPHLFGGFK@	748	Znrf2	E3 ubiquitin-protein ligase ZNRF2	1.8798

IPI:IPI00225062.2	R{circumflex over ( )}SSSELSPEVVEK@	749	Srrm2	Isoform	3 of Serine/arginine repetitive	1.8718
				matrix protein 2

IPI:IPI00309059.7	STSPIIGS*PPVR{circumflex over ( )}	750	Patl1	Protein PAT1 homolog 1	1.8673

IPI:IPI00751009.1	TSEFPTPLFSGPLEPVACGS	751	Srpk2	serine/arginine-rich protein specific	1.859
	VISEGSPLT*EQEESSPSHD			kinase 2
	R{circumflex over ( )}SR{circumflex over ( )}

IPI:IPI00808277.2	SAPASPNHAGVLSAHSSGA	752	Foxk2	Isoform 1 of Forkhead box protein K2	1.8546
	QTPESLSR{circumflex over ( )}EGS*PAPLEP
	EPGASQPK@

IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	753	Ndrg3	Protein NDRG3	1.8482

IPI:IPI00761759.1	SSSGSEHST*EGSVSLGDG	754	Larp4	Putative uncharacterized protein	1.8482
	PLSR{circumflex over ( )}

IPI:IPI00122594.4	EVSVSSVTEEPK@LDSSQ	755	Ahctf1	AT-hook-containing transcription	1.8472
	LPLQTGLDVPATPR{circumflex over ( )}			factor 1

IPI:IPI00122594.4	EVSVSSVTEEPK@LDSSQ	756	Ahctf1	AT-hook-containing transcription	1.8472
	LPLQTGLDVPATPR{circumflex over ( )}			factor 1

IPI:IPI00268673.5	AGTTVPESIHSFIGDGLVK	757	Mtor	Isoform	1 of FKBP12-rapamycin	1.8442
	@PEALNK@K@			complex-associated protein

IPI:IPI00314502.5	VHGLPTTS*PSGVNMAELA	758	Tcfeb	Transcription factor EB	1.8426
	QQVVK@

IPI:IPI00125319.1	TTS*FAESCK@PVQQPSAF	759	Gsk3b	Glycogen synthase kinase-3 beta	1.8417
	GSMK@

IPI:IPI00125319.1	TTS*FAESCK@PVQQPSAF	760	Gsk3b	Glycogen synthase kinase-3 beta	1.8417
	GSMK@

IPI:IPI00116285.2	SQDSYPVS*PR{circumflex over ( )}PFSSPSMS	761	Ranbp9	RAN binding protein 9	1.8399
	*PSHGMSIHSLAPGK@

IPI:IPI00323510.5	TDNSVASSPSSAISTATPS	762	Kdm6a	Isoform	1 of Lysine-specific	1.8307
	PK@			demethylase 6A

IPI:IPI00229859.1	GHPSAGAEEEGGSDGSA	763	Eif3b	Eif3b protein	1.826
	AEAEPR
PI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	764	Ndrg3	Protein NDRG3	1.8106

IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	765	Ndrg3	Protein NDRG3	1.8106

IPI:IPI00225062.2	GCSPPK@SPEK@PPQST	766	Srrm2	Isoform	3 of Serine/arginine repetitive	1.8106
	SSESCPPS*PQPTK@			matrix protein	2

IPI:IPI00405752.3	VLASSLSPYR{circumflex over ( )}EGR{circumflex over ( )}	767	C130092O11Rik	Isoform 1 of Uncharacterized protein	1.8087
				KIAA1680

IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	768	Ndrg3	Protein NDRG3	1.8041

IPI:IPI00130920.1	RSES*PFEGK	769	Mtap1b	Microtubule-associated protein 1B	1.7981

IPI:IPI00136107.1	THSTSSS*IGSGESPFSR{circumflex over ( )}	770	Ndrg3	Protein NDRG3	1.7967

IPI:IPI00336713.1	CSPVPGLSSSPSGSPLHG	771	Bcas3	Isoform	1 of Breast carcinoma-	1.7932
	K@			amplified sequence 3 homolog

IPI:IPI00323349.2	GYPPPIAAK@PAFGR{circumflex over ( )}PILK	772	Tjp2	Tight junction protein ZO-2	1.7869
	@PSTPVPMPESEEVGES
	TEEQEDAPR{circumflex over ( )}

IPI:IPI00225062.2	GCSPPK@SPEK@PPQST	773	Srrm2	Isoform	3 of Serine/arginine repetitive	1.7868
	SSESCPPS*PQPTK@			matrix protein 2

IPI:IPI00225062.2	GCSPPKSPEKPPQSTSSE	774	Srrm2	Isoform 3 of Serine/arginine repetitive	1.7868
	SCPPS*PQPTK			matrix protein 2

IPI:IPI00808277.2	SAPASPNHAGVLSAHSSGA	775	Foxk2	Isoform 1 of Forkhead box protein K2	1.7848
	QTPESLSR{circumflex over ( )}EGS*PAPLEP
	EPGASQPK@

IPI:IPI00136107.1	THSTSSS*IGSGESPFSR{circumflex over ( )}	776	Ndrg3	Protein NDRG3	1.7706

IPI:IPI00107958.1	TSDIFGS*PVTATAPLAHPN	777	Hn1l	Hematological and neurological	1.7561
	K@PK@			expressed 1-like protein

IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	778	Ndrg3	Protein NDRG3	1.7555

IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	779	Ndrg3	Protein NDRG3	1.7525

IPI:IPI00130920.1	RSES*PFEGK	780	Mtap1b	Microtubule-associated protein 1B	1.7508

IPI:IPI00130920.1	RSES*PFEGK	781	Mtap1b	Microtubule-associated protein 1B	1.7508

IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	782	Ndrg3	Protein NDRG3	1.7499

IPI:IPI00136107.1	THSTSSSIGSGESPFSR	783	Ndrg3	Protein NDRG3	1.7499

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	784	Eif4ebp1	Eukaryotic translation initiation	1.7456
	GGTLFS*TTPGGTRIIYDR			factor 4E-binding protein 1

IPI:IPI00468516.3	R{circumflex over ( )}LAAQES*SEAEDVTVDR{circumflex over ( )}	785	D6Wsu116e	Isoform	1 of Protein FAM21	1.7446
	GPVAQLSSS*PVLPNGHQP
	LLQPR{circumflex over ( )}

IPI:IPI00137501.1	TEMDKSPFNSPSPQDS*PR	786	Nfic	Isoform	1 of Nuclear factor 1 C-type	1.7419

IPI:IPI00225062.2	GCSPPK@SPEK@PPQS*	787	Srrm2	Isoform 3 of Serine/arginine repetitive	1.7403
	TSSESCPPSPQPTK@			matrix protein 2

IPI:IPI00225062.2	GCSPPK@SPEK@PPQST	788	Srrm2	Isoform	3 of Serine/arginine repetitive	1.7403
	SSESCPPS*PQPTK@			matrix protein 2

IPI:IPI00225062.2	GCSPPKSPEKPPQSTSS*	789	Srrm2	Isoform	3 of Serine/arginine repetitive	1.7403
	ESCPPSPQPTK			matrix protein 2

IPI:IPI00336973.2	GLNLDGTPALSTLGGFS*PA	790	Ccnl1	Isoform	1 of Cyclin-L1	1.7368
	SK@PS*SPR{circumflex over ( )}

IPI:IPI00107958.1	TSDIFGS*PVTATAPLAHPN	791	Hn1l	Hematological and neurological	1.7353
	K@PK@			expressed 1-like protein

IPI:IPI00227209.3	R{circumflex over ( )}LSSTSLASGHSVR{circumflex over ( )}	792	Prkd2	Serine/threonine-protein kinase D2	1.7328

IPI:IPI00225062.2	RSSSELSPEVVEK	793	Srrm2	Isoform 3 of Serine/arginine repetitive	1.7324
				matrix protein 2

IPI:IPI00225062.2	R{circumflex over ( )}SSSELSPEVVEK@	794	Srrm2	Isoform	3 of Serine/arginine repetitive	1.7324
				matrix protein 2

IPI:IPI00225062.2	RSSSELSPEVVEK	795	Srrm2	Isoform	3 of Serine/arginine repetitive	1.7324
				matrix protein 2

IPI:IPI00225062.2	R{circumflex over ( )}SSSELSPEVVEK@	796	Srrm2	Isoform	3 of Serine/arginine repetitive	1.7324
				matrix protein 2

IPI:IPI00929779.1	S*EGLSQEATPSQDLIQHSC	797	Usp36	Ubiquitin specific peptidase 36	1.7192
	SPVDHSEPEAR{circumflex over ( )}

IPI:IPI00467930.2	LVIGSLPAHLSPHLFGGFK	798	Znrf2	E3 ubiquitin-protein ligase ZNRF2	1.7137

IPI:IPI00467930.2	LVIGSLPAHLSPHLFG	799	Znrf2	E3 ubiquitin-protein ligase ZNRF2	1.7137
	GFK@

IPI:IPI00459443.5	DLQSEFGVATDSHHSSFGS	800	Tnks1bp1	182 kDa tankyrase-1-binding protein	1.7011
	SSWSQDTSQNYSLGGR{circumflex over ( )}S*
	PVGDTGLGK@

IPI:IPI00785400.1	LTSHTPGLDDEKEAS*ENET	801	Syne2	Syne2 protein	1.7003
	*DIEDPREIQADSWR

IPI:IPI00136107.1	THST*SSSIGSGESPFSR{circumflex over ( )}	802	Ndrg3	Protein NDRG3	1.6838

IPI:IPI00136107.1	THST*SSSIGSGESPFSR{circumflex over ( )}	803	Ndrg3	Protein NDRG3	1.6838

IPI:IPI00336713.1	QGGRCSPVPGLSSSPSG	804	Bcas3	Isoform	1 of Breast carcinoma-	1.6818
	SPLHGK			amplified sequence 3 homolog

IPI:IPI00336713.1	QGGR{circumflex over ( )}CSPVPGLSSSPSG	805	Bcas3	Isoform	1 of Breast carcinoma-	1.6818
	SPLHGK@			amplified sequence 3 homolog

IPI:IPI00380354.1	LQES*PKLSQANGTR	806	Llgl1	lethal giant larvae homolog 1 isoform 1	1.6806

IPI:IPI00128904.1	VMTIPYQPMPASS*PVICAG	807	Pcbp1	Poly(rC)-binding protein 1	1.6782
	GQDR

IPI:IPI00380107.4	LVIPSATTKSPPEITVT	808	Cabin1	calcineurin binding protein 1	1.6782
	PPT*PTLLSPK

IPI:IPI00153375.1	SACFSPVSLSPR{circumflex over ( )}PCS*PF	809	Pdlim2	PDZ and LIM domain protein 2	1.6659
	STPPPTSPVALSK@

IPI:IPI00467930.2	LVIGS*LPAHLSPHLFGGFK@	810	Znrf2	E3 ubiquitin-protein ligase ZNRF2	1.6655

IPI:IPI00467930.2	LVIGS*LPAHLSPHLFGGFK@	811	Znrf2	E3 ubiquitin-protein ligase ZNRF2	1.6655

IPI:IPI00221581.1	SR{circumflex over ( )}TGSESS*QTGASATS	812	Eif4b	Eukaryotic translation initiation	1.6642
	GR{circumflex over ( )}			factor 4B

IPI:IPI00318938.6	NSPVAK@TPPK@DLPAIP	813	Eif4ebp1	Eukaryotic translation initiation	1.6633
	GVTSPTSDEPPMQASQSQL			factor 4E-binding protein 1
	PSSPEDK@

IPI:IPI00120529.2	NK@PLS*PIK@LTPTSVLDY	814	Rfc1	Rfc1 protein	1.6568
	FGTESVQR{circumflex over ( )}

IPI:IPI00225062.2	RSSSELSPEVVEK	815	Srrm2	Isoform 3 of Serine/arginine repetitive	1.6324
				matrix protein 2

IPI:IPI00225062.2	R{circumflex over ( )}SSSELSPEVVEK@	816	Srrm2	Isoform	3 of Serine/arginine repetitive	1.6324
				matrix protein 2

IPI:IPI00225062.2	R{circumflex over ( )}SSSELSPEVVEK@	817	Srrm2	Isoform 3 of Serine/arginine repetitive	1.6324
				matrix protein 2

IPI:IPI00225062.2	RSSSELSPEVVEK	818	Srrm2	Isoform 3 of Serine/arginine repetitive	1.6324
				matrix protein 2

IPI:IPI00225062.2	R{circumflex over ( )}SSSELSPEVVEK@	819	Srrm2	Isoform	3 of Serine/arginine repetitive	1.6324
				matrix protein 2

IPI:IPI00225062.2	R{circumflex over ( )}SSSELSPEVVEK@	820	Srrm2	Isoform	3 of Serine/arginine repetitive	1.6324
				matrix protein 2

IPI:IPI00761759.1	SSSGSEHSTEGS*VSLGDG	821	Larp4	Putative uncharacterized protein	1.6294
	PLSR{circumflex over ( )}

IPI:IPI00309059.7	R{circumflex over ( )}STSPIIGS*PPVR{circumflex over ( )}	822	Patl1	Protein PAT1 homolog 1	1.6275

IPI:IPI00309059.7	R{circumflex over ( )}STSPIIGS*PPVR{circumflex over ( )}	823	Patl1	Protein PAT1 homolog 1	1.6245

IPI:IPI00317401.6	AESPETSAVESTQSTPQK	824	Pds5b	Isoform	1 of Sister chromatid cohesion	1.623
	@GR{circumflex over ( )}			protein PDS5 homolog B

IPI:IPI00317401.6	AESPETSAVESTQSTPQK	825	Pds5b	Isoform	1 of Sister chromatid cohesion	1.623
	@GR{circumflex over ( )}			protein PDS5 homolog B

IPI:IPI00115620.1	AS*LYNAVTTEDVEK	826	Psat1	Phosphoserine aminotransferase	1.6216

IPI:IPI00126124.2	TFSECSYPETEEEAEALP	827	St5	Isoform	1 of Suppression of	1.6159
	GR{circumflex over ( )}			tumorigenicity 5

IPI:IPI00421179.1	SFS*K@EVEER{circumflex over ( )}	828	Eif4g1	Isoform	1 of Eukaryotic translation	1.6081
				initiation factor 4 gamma 1

IPI:IPI00225062.2	GCSPPK@SPEK@PPQST	829	Srrm2	Isoform	3 of Serine/arginine repetitive	1.608
	SSESCPPS*PQPTK@			matrix protein	2

IPI:IPI00227934.3	AAPSSEGDSCDGVEATDA	830	Cux1	cut-like homeobox 1 isoform a	1.6068
	EEPGGNIVATK@

IPI:IPI00314240.5	K@LEK@EEEEGISQESS*	831	Hmga1	Isoform HMG-I of High mobility group	1.6058
	EEEQ			protein HMG-I/HMG-Y

IPI:IPI00111754.6	LDT*FCGSPPYAAPELFQGK	832	Mark2	Isoform 2 of Serine/threonine-protein	1.5994
				kinase MARK2

IPI:IPI00125319.1	TTS*FAESCK@PVQQPSAF	833	Gsk3b	Glycogen synthase kinase-3 beta	1.5966
	GSMK@

IPI:IPI00320905.7	LGEQGPEPGPTPPQTPTP	834	Arhgap17	Isoform 1 of Rho GTPase-activating	1.5946
	PS*TPPLAK			protein 17

IPI:IPI00380737.1	TSSVSSLASACTGGIPSSS	835	Pom121	Nuclear envelope pore membrane	1.5883
	R{circumflex over ( )}			protein POM 121

IPI:IPI00122594.4	TTPLASPSLSPGR{circumflex over ( )}	836	Ahctf1	AT-hook-containing transcription	1.5845
				factor 1

IPI:IPI00125319.1	TTS*FAESCK@PVQQPSAF	837	Gsk3b	Glycogen synthase kinase-3 beta	1.5821
	GSMK@

IPI:IPI00381019.1	KRSPSPSPTPEAK	838	Smarcc2	Isoform	2 of SWI/SNF complex subunit	1.5795
				SMARCC2

IPI:IPI00111754.6	LDT*FCGSPPYAAPELFQGK@	839	Mark2	Isoform 2 of Serine/threonine-protein	1.578
				kinase MARK2

IPI:IPI00111754.6	LDT*FCGSPPYAAPELFQGK@	840	Mark2	Isoform 2 of Serine/threonine-protein	1.578
				kinase MARK2

IPI:IPI00136107.1	THS*TSSSIGSGESPFSR{circumflex over ( )}	841	Ndrg3	Protein NDRG3	1.576

IPI:IPI00136107.1	THS*TSSSIGSGESPFSR{circumflex over ( )}	842	Ndrg3	Protein NDRG3	1.576

IPI:IPI00128904.1	VMTIPYQPMPASS*PVICAG	843	Pcbp1	Poly(rC)-binding protein 1	1.5752
	GQDR

IPI:IPI00225062.2	SSSPVTELTAR{circumflex over ( )}	844	Srrm2	Isoform	3 of Serine/arginine repetitive	1.5701
				matrix protein 2

IPI:IPI00421179.1	SFS*KEVEER	845	Eif4g1	Isoform 1 of Eukaryotic translation	1.5642
				initiation factor 4 gamma 1

IPI:IPI00421179.1	SFS*K@EVEER{circumflex over ( )}	846	Eif4g1	Isoform 1 of Eukaryotic translation	1.5642
				initiation factor 4 gamma 1

IPI:IPI00421179.1	SFS*KEVEER	847	Eif4g1	Isoform 1 of Eukaryotic translation	1.5642
				initiation factor 4 gamma 1

IPI:IPI00421179.1	SFS*K@EVEER{circumflex over ( )}	848	Eif4g1	Isoform 1 of Eukaryotic translation	1.5642
				initiation factor 4 gamma 1

IPI:IPI00115492.1	HSLSSESQAPEDIAPPGS*S	849	Eps8l2	Isoform 1 of Epidermal growth factor	1.564
	PHANR{circumflex over ( )}			receptor kinase substrate 8-like
				protein
2

IPI:IPI00761759.1	SSSGS*EHSTEGSVSLGD	850	Larp4	Putative uncharacterized protein	1.5621
	GPLSR{circumflex over ( )}

IPI:IPI00136107.1	THSTS*SSIGSGESPFSR{circumflex over ( )}	851	Ndrg3	Protein NDRG3	1.5584

IPI:IPI00111754.6	LDT*FCGSPPYAAPELFQGK	852	Mark2	Isoform	2 of Serine/threonine-protein	1.5403
				kinase MARK2

IPI:IPI00336973.2	GLNLDGTPALSTLGGFSP	853	Ccnl1	Isoform	1 of Cyclin-L1	1.5363
	ASK@PS*SPR{circumflex over ( )}

IPI:IPI00229739.4	LVVSSPTSPK@GK@	854	Plekhm1	Pleckstrin homology domain-containing	1.5286
				family M member 1

IPI:IPI00126124.2	TFSECSYPETEEEAEALP	855	St5	Isoform	1 of Suppression of	1.526
	GR{circumflex over ( )}			tumorigenicity 5

IPI:IPI00122594.4	TTPLASPSLSPGR{circumflex over ( )}	856	Ahctf1	AT-hook-containing transcription	1.5177
				factor 1

IPI:IPI00229739.4	LWSSPTSPKGK	857	Plekhm1	Pleckstrin homology domain-containing	1.515
				family M member 1

IPI:IPI00421179.1	SFS*K@EVEER{circumflex over ( )}	858	Eif4g1	Isoform 1 of Eukaryotic translation	1.5113
				initiation factor 4 gamma 1

IPI:IPI00421179.1	SFS*KEVEER	859	Eif4g1	Isoform 1 of Eukaryotic translation	1.5113
				initiation factor 4 gamma 1

IPI:IPI00349069.4	LFGT*SPAAEVTPSPPEPAP	860	B230208H17Rik	Putative GTP-binding protein Parf	1.5073
	ALEAPAR{circumflex over ( )}

IPI:IPI00605037.1	KETESEAEDDNLDDLER	861	Srrm1	Isoform 1 of Serine/arginine repetitive	1.5038
				matrix protein 1

IPI:IPI00381244.8	KEELGASS*PGYGPPNLGC	862	Mll2	similar to myeloid/lymphoid or mixed-	1.4993
	VDS*PSAGPHLGGLELK			lineage leukemia 2

IPI:IPI00421179.1	SFS*K@EVEER{circumflex over ( )}	863	Eif4g1	Isoform 1 of Eukaryotic translation	1.4913
				initiation factor 4 gamma 1

IPI:IPI00421179.1	SFS*KEVEER	864	Eif4g1	Isoform 1 of Eukaryotic translation	1.4913
				initiation factor 4 gamma 1

IPI:IPI00421179.1	SFS*KEVEER	865	Eif4g1	Isoform 1 of Eukaryotic translation	1.4913
				initiation factor 4 gamma 1

IPI:IPI00421179.1	SFS*K@EVEER{circumflex over ( )}	866	Eif4g1	Isoform 1 of Eukaryotic translation	1.4913
				initiation factor 4 gamma 1

IPI:IPI00136572.4	YEENPAWPGTSTHNGPNG	867	Zc3hav1	Isoform 1 of Zinc finger CCCH-type	1.4864
	FSQIMDET*PNVSK@SSPT			antiviral protein 1
	GFGIK@

IPI:IPI00318938.6	NSPVAK@TPPK@DLPAIP	868	Eif4ebp1	Eukaryotic translation initiation	1.4819
	GVTSPTSDEPPMQASQSQL			factor 4E-binding protein 1
	PSSPEDK@

IPI:IPI00318938.6	NSPVAK@TPPK@DLPAIP	869	Eif4ebp1	Eukaryotic translation initiation	1.4819
	GVTSPTSDEPPMQASQSQL			factor 4E-binding protein 1
	PSSPEDK@

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	870	Eif4ebp1	Eukaryotic translation initiation	1.4819
	SPTSDEPPMQASQSQLPSS			factor 4E-binding protein 1
	PEDK

IPI:IPI00318938.6	NSPVAK@TPPK@DLPAIP	871	Eif4ebp1	Eukaryotic translation initiation	1.4819
	GVTSPTSDEPPMQASQSQL			factor 4E-binding protein 1
	PSSPEDK@

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	872	Eif4ebp1	Eukaryotic translation initiation	1.4819
	SPTSDEPPMQASQSQLPSS			factor 4E-binding protein 1
	PEDK

IPI:IPI00228948.3	S*FGDK@DLILPNGGTPAG	873	Snx30	Sorting nexin-30	1.4774
	TASPASSSSLLNR{circumflex over ( )}

IPI:IPI00330246.2	WAAHGTS*PEDFSLTLGAR{circumflex over ( )}	874	Phldb1	Isoform	2 of Pleckstrin homology-like	1.474
				domain family B member 1

IPI:IPI00229739.4	LVVSSPTSPK@GK@	875	Plekhm1	Pleckstrin homology domain-containing	1.4728
				family M member 1

IPI:IPI00229739.4	LVVSSPTSPKGK	876	Plekhm1	Pleckstrin homology domain-containing	1.4728
				family M member 1

IPI:IPI00453603.1	FIGSPR{circumflex over ( )}TPVS*PVK@FSP	877	Rps6kb1	Isoform Alpha I of Ribosomal protein	1.4727
	GDFWGR{circumflex over ( )}			S6 kinase beta-1

IPI:IPI00136107.1	THS*TSSSIGSGESPFSR{circumflex over ( )}	878	Ndrg3	Protein NDRG3	1.4724

IPI:IPI00136107.1	THS*TSSSIGSGESPFSR{circumflex over ( )}	879	Ndrg3	Protein NDRG3	1.4724

IPI:IPI00317401.6	AESPETSAVESTQSTPQK	880	Pds5b	Isoform	1 of Sister chromatid cohesion	1.4711
	@GR{circumflex over ( )}			protein PDS5 homolog B

IPI:IPI00317401.6	AESPETSAVESTQSTPQK	881	Pds5b	Isoform	1 of Sister chromatid cohesion	1.4711
	@GR{circumflex over ( )}			protein PDS5 homolog B

IPI:IPI00453603.1	FIGSPRTPVS*PVKFSPGD	882	Rps6kb1	Isoform Alpha I of Ribosomal protein	1.4689
	FWGR			S6 kinase beta-1

IPI:IPI00453603.1	FIGSPR{circumflex over ( )}TPVS*PVK@FSP	883	Rps6kb1	Isoform Alpha I of Ribosomal protein	1.4689
	GDFWGR{circumflex over ( )}			S6 kinase beta-1

IPI:IPI00453603.1	FIGSPR{circumflex over ( )}TPVS*PVK@FSP	884	Rps6kb1	Isoform Alpha I of Ribosomal protein	1.4689
	GDFWGR{circumflex over ( )}			S6 kinase beta-1

IPI:IPI00380737.1	TSSVSSLASACTGGIPSSS	885	Pom121	Nuclear envelope pore membrane	1.4684
	R{circumflex over ( )}			protein POM 121

IPI:IPI00380737.1	TSSVSSLASACTGGIPSSS	886	Pom121	Nuclear envelope pore membrane	1.4684
	R{circumflex over ( )}			protein POM 121

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	887	Eif4ebp1	Eukaryotic translation initiation	1.4676
	SPTSDEPPMQASQSQLPSS			factor 4E-binding protein 1
	PEDK

IPI:IPI00317401.6	AESPETSAVESTQSTPQK	888	Pds5b	Isoform	1 of Sister chromatid cohesion	1.4613
	@GR{circumflex over ( )}			protein PDS5 homolog B

IPI:IPI00317401.6	AESPETSAVESTQSTPQK	889	Pds5b	Isoform	1 of Sister chromatid cohesion	1.4526
	@GR{circumflex over ( )}			protein PDS5 homolog B

IPI:IPI00317401.6	AESPETSAVESTQSTPQK	890	Pds5b	Isoform 1 of Sister chromatid cohesion	1.4526
	@GR{circumflex over ( )}			protein PDS5 homolog B

IPI:IPI00136107.1	SRTHSTSSSIGSGESPFSR	891	Ndrg3	Protein NDRG3	1.451

IPI:IPI00118438.4	KPPAPPSPVQSQSPST*N	892	Srrm1	Isoform	2 of Serine/arginine repetitive	1.45
	WS*PAVPAKK			matrix protein 1

IPI:IPI00123410.5	TIS*AQDTLAYATALLNEK@	893	Usp24	Isoform	1 of Ubiquitin carboxyl-	1.4486
				terminal hydrolase 24

IPI:IPI00411026.1	SLHPWYGITPTSS*PK@	894	Micall1	Isoform 1 of MICAL-like protein 1	1.4479

IPI:IPI00761759.1	SSSGSEHST*EGSVSLGD	895	Larp4	Putative uncharacterized protein	1.4347
	GPLSR{circumflex over ( )}

IPI:IPI00313307.3	SYQNSPSSEDGIR{circumflex over ( )}PLPEY	896	Med1	Isoform 4 of Mediator of RNA	1.4214
	STEK@			polymerase II transcription subunit 1

IPI:IPI00313307.3	SYQNSPSSEDGIR{circumflex over ( )}PLPEY	897	Med1	Isoform	4 of Mediator of RNA	1.4158
	STEK@			polymerase II transcription subunit 1

IPI:IPI00129264.1	ATSR{circumflex over ( )}PINLGPSS*PNTEIHW	898	Sorbs3	Vinexin	1.4122
	TPYR{circumflex over ( )}

IPI:IPI00330773.3	ASYSGTSPSHSFISGEPD	899	Phldb2	Isoform	1 of Pleckstrin homology-like	1.403
	R{circumflex over ( )}			domain family B member 2

IPI:IPI00116442.1	TR{circumflex over ( )}LASESANDDNEDS	900	Hdgfrp2	Isoform	3 of Hepatoma-derived growth	1.3985
				factor-related protein 2

IPI:IPI00309059.7	STSPIIGSPPVR{circumflex over ( )}	901	Patl1	Protein PAT1 homolog 1	1.3955

IPI:IPI00309059.7	STSPIIGSPPVR{circumflex over ( )}	902	Patl1	Protein PAT1 homolog 1	1.3937

IPI:IPI00309059.7	STSPIIGSPPVR{circumflex over ( )}	903	Patl1	Protein PAT1 homolog 1	1.3937

IPI:IPI00309059.7	STSPIIGSPPVR	904	Patl1	Protein PAT1 homolog 1	1.3937

IPI:IPI00318938.6	NS*PVAK@TPPK@DLPAIP	905	Eif4ebp1	Eukaryotic translation initiation	1.3906
	GVTSPTS*DEPPMQASQSQ			factor 4E-binding protein 1
	LPSSPEDK@

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	906	Eif4ebp1	Eukaryotic translation initiation	1.3906
	SPTSDEPPMQASQSQLPSS			factor 4E-binding protein 1
	PEDK

IPI:IPI00108388.1	R{circumflex over ( )}PYTGNPQYTYNNWS*PP	907	Usp9x	Probable ubiquitin carboxyl-terminal	1.3905
	VQSNETSNGYFLER{circumflex over ( )}			hydrolase FAF-X

IPI:IPI00108388.1	RPYTGNPQYTYNNWS*PPV	908	Usp9x	Probable ubiquitin carboxyl-terminal	1.3905
	QSNETSNGYFLER			hydrolase FAF-X

IPI:IPI00309059.7	STSPIIGSPPVR{circumflex over ( )}	909	Patl1	Protein PAT1 homolog 1	1.3897

IPI:IPI00309059.7	STSPIIGSPPVR{circumflex over ( )}	910	Patl1	Protein PAT1 homolog 1	1.3897

IPI:IPI00556837.1	DTVIIVSEPSEDEESHDLP	911	Smarcad1	Isoform 1 of SWI/SNF-related matrix-	1.3868
	SVTR			associated actin-dependent regulator of
				chromatin subfamily A containing
				DEAD/H box 1

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	912	Eif4ebp1	Eukaryotic translation initiation	1.3764
	SPTSDEPPMQASQSQLPSS			factor 4E-binding protein 1
	PEDK

IPI:IPI00467843.2	GGGASSPAPVVFTVGS*PP	913	Ulk1	Putative uncharacterized protein	1.3745
	SGAT*PPQSTR{circumflex over ( )}

IPI:IPI00467843.2	GGGASSPAPVVFTVGS*PP	914	Ulk1	Putative uncharacterized protein	1.3745
	SGATPPQST*R{circumflex over ( )}

IPI:IPI00467843.2	GGGASSPAPVVFTVGS*PP	915	Ulk1	Putative uncharacterized protein	1.3702
	SGAT*PPQSTR{circumflex over ( )}

IPI:IPI00330773.3	ASYSGTS*PSHSFISGEPDR{circumflex over ( )}	916	Phldb2	Isoform	1 of Pleckstrin homology-like	1.3697
				domain family B member 2

IPI:IPI00330773.3	AS*YSGTSPSHSFISGEPDR	917	Phldb2	Isoform	1 of Pleckstrin homology-like	1.3697
				domain family B member 2

IPI:IPI00330773.3	ASYSGTS*PSHSFISGEPDR{circumflex over ( )}	918	Phldb2	Isoform	1 of Pleckstrin homology-like	1.3697
				domain family B member 2

IPI:IPI00117932.2	SPPVQPHTPVTIS*LGTAPS	919	Sin3a	Isoform 1 of Paired amphipathic helix	1.3566
	LQNNQPVEFNHAINYVNK@			protein Sin3a

IPI:IPI00309059.7	STSPIIGSPPVR{circumflex over ( )}	920	Patl1	Protein PAT1 homolog 1	1.3334

IPI:IPI00309059.7	STSPIIGSPPVR{circumflex over ( )}	921	Patl1	Protein PAT1 homolog 1	1.3334

IPI:IPI00224153.6	VHHSVQTFQEDSLPVAHS	922	Ptpn21	Tyrosine-protein phosphatase non-	1.3332
	LQEVSEPLTAAR{circumflex over ( )}			receptor type 21

IPI:IPI00224153.6	VHHSVQTFQEDSLPVAHS	923	Ptpn21	Tyrosine-protein phosphatase non-	1.3332
	LQEVSEPLTAAR			receptor type 21

IPI:IPI00136572.4	FHHNSLEVLSTVSPLGSG	924	Zc3hav1	Isoform	1 of Zinc finger CCCH-type	1.3312
	PPS*PDVTGCK@DPLEDVS			antiviral protein 1
	ADVTQK@

IPI:IPI00309059.7	STSPIIGSPPVR	925	Patl1	Protein PAT1 homolog 1	1.3311

IPI:IPI00309059.7	STSPIIGSPPVR{circumflex over ( )}	926	Patl1	Protein PAT1 homolog 1	1.3311

IPI:IPI00309059.7	STSPIIGSPPVR{circumflex over ( )}	927	Patl1	Protein PAT1 homolog 1	1.3311

IPI:IPI00309059.7	STSPIIGSPPVR	928	Patl1	Protein PAT1 homolog 1	1.3311

IPI:IPI00309059.7	R{circumflex over ( )}STSPIIGSPPVR{circumflex over ( )}	929	Patl1	Protein PAT1 homolog 1	1.3267

IPI:IPI00229739.4	LVVSSPTSPK@	930	Plekhm1	Pleckstrin homology domain-containing	1.3252
				family M member 1

IPI:IPI00321774.2	HSSIS*PVR{circumflex over ( )}LPLNSSLGAEL	931	Crkrs	Isoform	2 of Cell division cycle 2-	1.3231
	SR{circumflex over ( )}			related protein kinase 7

IPI:IPI00129264.1	ATSR{circumflex over ( )}PINLGPS*SPNTEIHW	932	Sorbs3	Vinexin	1.3226
	TPYR{circumflex over ( )}

IPI:IPI00380354.1	LQES*PKLSQANGTR	933	Llgl1	lethal giant larvae homolog 1 isoform 1	1.3225

IPI:IPI00320905.7	LGEQGPEPGPTPPQTPT*	934	Arhgap17	Isoform 1 of Rho GTPase-activating	1.3166
	PPST*PPLAK			protein	17

IPI:IPI00381495.5	TVSSPIPYTPSPSSSR{circumflex over ( )}PIS	935	Ccdc6	coiled-coil domain containing 6	1.3165
	PGLSYASHTVGFTPPTSL
	TR{circumflex over ( )}

IPI:IPI00330773.3	ASYSGTSPSHSFISGEPD	936	Phldb2	Isoform	1 of Pleckstrin homology-like	1.3149
	R{circumflex over ( )}			domain family B member 2

IPI:IPI00660767.2	VDTAASSSWLAGSCSPVS	937	Ibtk	Isoform 2 of Inhibitor of Bruton	1.3143
	PPVVDLR{circumflex over ( )}			tyrosine kinase

IPI:IPI00309059.7	RSTSPIIGSPPVR	938	Patl1	Protein PAT1 homolog 1	1.3126

IPI:IPI00309059.7	R{circumflex over ( )}STSPIIGSPPVR{circumflex over ( )}	939	Patl1	Protein PAT1 homolog 1	1.3126

IPI:IPI00309059.7	R{circumflex over ( )}STSPIIGSPPVR{circumflex over ( )}	940	Patl1	Protein PAT1 homolog 1	1.3126

IPI:IPI00309059.7	RSTSPIIGSPPVR	941	Patl1	Protein PAT1 homolog 1	1.3123

IPI:IPI00309059.7	RSTSPIIGSPPVR	942	Patl1	Protein PAT1 homolog 1	1.3122

IPI:IPI00309059.7	R{circumflex over ( )}STSPIIGSPPVR{circumflex over ( )}	943	Patl1	Protein PAT1 homolog 1	1.3122

IPI:IPI00309059.7	R{circumflex over ( )}STSPIIGSPPVR{circumflex over ( )}	944	Patl1	Protein PAT1 homolog 1	1.3122

IPI:IPI00112593.2	STSSPK@	945	Spert	Isoform 2 of Spermatid-associated	1.3085
				protein

IPI:IPI00129264.1	ATSR{circumflex over ( )}PINLGPSS*PNTEIHW	946	Sorbs3	Vinexin	1.3056
	TPYR{circumflex over ( )}

IPI:IPI00129264.1	ATSRPINLGPSS*PNTEIHW	947	Sorbs3	Vinexin	1.3056
	TPYR

IPI:IPI00116442.1	TR{circumflex over ( )}LASESANDDNEDS	948	Hdgfrp2	Isoform	3 of Hepatoma-derived growth	1.3036
				factor-related protein 2

IPI:IPI00313307.3	SYQNSPSSEDGIRPLPEY	949	Med1	Isoform 4 of Mediator of RNA	1.298
	STEK			polymerase II transcription subunit 1

IPI:IPI00313307.3	SYQNSPSSEDGIR{circumflex over ( )}PLPEY	950	Med1	Isoform 4 of Mediator of RNA	1.298
	STEK@			polymerase II transcription subunit 1

IPI:IPI00313307.3	SYQNSPSSEDGIR{circumflex over ( )}PLPEY	951	Med1	Isoform	4 of Mediator of RNA	1.298
	STEK@			polymerase II transcription subunit 1

IPI:IPI00330773.3	ASYSGTS*PSHSFISGEPDR{circumflex over ( )}	952	Phldb2	Isoform	1 of Pleckstrin homology-like	1.2944
				domain family B member 2

IPI:IPI00330773.3	ASYSGTS*PSHSFISGEPDR	953	Phldb2	Isoform	1 of Pleckstrin homology-like	1.2944
				domain family B member 2

IPI:IPI00330773.3	ASYSGTS*PSHSFISGEPDR{circumflex over ( )}	954	Phldb2	Isoform	1 of Pleckstrin homology-like	1.2944
				domain family B member 2

IPI:IPI00153986.2	GTSR{circumflex over ( )}PGTPSAEAASTSS	955	Gtf2f1	General transcription factor IIF	1.2876
	TLR{circumflex over ( )}			subunit 1

IPI:IPI00153986.2	GTSRPGTPSAEAASTSST	956	Gtf2f1	General transcription factor IIF	1.2876
	LR			subunit	1

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	957	Eif4ebp1	Eukaryotic translation initiation	1.2862
	GGTLFS*TTPGGTR{circumflex over ( )}IIYDR{circumflex over ( )}			factor 4E-binding protein 1
	K@

IPI:IPI00221581.1	SR{circumflex over ( )}TGS*ESSQTGASATSG	958	Eif4b	Eukaryotic translation initiation	1.2845
	R{circumflex over ( )}			factor 4B

IPI:IPI00454104.1	TTEAPCSPGSQQPPSPDE	959	Scrib	Isoform 1 of Protein LAP4	1.2842
	LPANVK@

IPI:IPI00454104.1	TTEAPCSPGSQQPPSPDE	960	Scrib	Isoform	1 of Protein LAP4	1.2842
	LPANVK

IPI:IPI00454104.1	TTEAPCSPGSQQPPSPDE	961	Scrib	Isoform 1 of Protein LAP4	1.2842
	LPANVK

IPI:IPI00320905.7	LGEQGPEPGPTPPQTPTP	962	Arhgap17	Isoform 1 of Rho GTPase-activating	1.2828
	PST*PPLAK@			protein 17

IPI:IPI00381495.5	TVSSPIPYTPSPSSSR{circumflex over ( )}PIS	963	Ccdc6	coiled-coil domain containing 6	1.2808
	PGLSYASHTVGFTPPTSL
	TR{circumflex over ( )}

IPI:IPI00381495.5	TVSSPIPYTPSPSS*SR	964	Ccdc6	coiled-coil domain containing 6	1.2808
	PISPGLSYASHTVGFT*
	PPTSLTR

IPI:IPI00381495.5	TVSSPIPYTPSPSSSRPIS	965	Ccdc6	coiled-coil domain containing 6	1.2808
	PGLSYASHTVGFTPPTSL
	TR

IPI:IPI00309059.7	RSTSPIIGSPPVR	966	Patl1	Protein PAT1 homolog 1	1.2745

IPI:IPI00336973.2	GLNLDGTPALSTLGGFSP	967	Ccnl1	Isoform 1 of Cyclin-L1	1.273
	ASK@PS*SPR{circumflex over ( )}

IPI:IPI00336973.2	GLNLDGTPALSTLGGFSP	968	Ccnl1	Isoform	1 of Cyclin-L1	1.273
	ASKPSS*PR

IPI:IPI00309059.7	R{circumflex over ( )}STSPIIGSPPVR{circumflex over ( )}	969	Patl1	Protein PAT1 homolog 1	1.2725

IPI:IPI00309059.7	R{circumflex over ( )}STSPIIGSPPVR{circumflex over ( )}	970	Patl1	Protein PAT1 homolog 1	1.2725

IPI:IPI00309059.7	RSTSPIIGSPPVR	971	Patl1	Protein PAT1 homolog 1	1.2725

IPI:IPI00309059.7	R{circumflex over ( )}STSPIIGSPPVR{circumflex over ( )}	972	Patl1	Protein PAT1 homolog 1	1.2723

IPI:IPI00309059.7	RSTSPIIGSPPVR	973	Patl1	Protein PAT1 homolog 1	1.2723

IPI:IPI00309059.7	R{circumflex over ( )}STSPIIGSPPVR{circumflex over ( )}	974	Patl1	Protein PAT1 homolog 1	1.2723

IPI:IPI00309059.7	RSTSPIIGSPPVR	975	Patl1	Protein PAT1 homolog 1	1.2723

IPI:IPI00129264.1	ATSR{circumflex over ( )}PINLGPS*SPNTEIHW	976	Sorbs3	Vinexin	1.2719
	TPYR{circumflex over ( )}

IPI:IPI00318938.6	NSPVAK@TPPK@DLPAIP	977	Eif4ebp1	Eukaryotic translation initiation	1.2719
	GVTSPTSDEPPM#QASQSQ			factor 4E-binding protein 1
	LPSSPEDK@

IPI:IPI00115492.1	HSLSSESQAPEDIAPPGS*S	978	Eps8l2	Isoform	1 of Epidermal growth factor	1.2678
	PHANR{circumflex over ( )}			receptor kinase substrate 8-like
				protein
2

IPI:IPI00381495.5	TVSSPIPYTPSPSSSR{circumflex over ( )}PIS	979	Ccdc6	coiled-coil domain containing 6	1.2638
	PGLSYASHTVGFTPPTSL
	TR{circumflex over ( )}

IPI:IPI00381495.5	TVSSPIPYTPSPSSSRP	980	Ccdc6	coiled-coil domain containing 6	1.2638
	ISPGLSYASHTVGFT
	PPTSLTR

IPI:IPI00381495.5	TVSSPIPYTPSPSSSRP	981	Ccdc6	coiled-coil domain containing 6	1.2638
	ISPGLSYASHTVGFT
	PPTSLTR

IPI:IPI00381495.5	TVSSPIPYTPSPSSSR{circumflex over ( )}PIS	982	Ccdc6	coiled-coil domain containing 6	1.2638
	PGLSYASHTVGFTPPTSL
	TR{circumflex over ( )}

IPI:IPI00133917.1	YAQGFLPEK@PPQQDHTT*	983	Phc2	Isoform	1 of Polyhomeotic-like	1.2624
	TTDSEMEEPYLQESK@EE			protein	2
	GTPLK@

IPI:IPI00227209.3	R{circumflex over ( )}LSSTSLASGHSVR{circumflex over ( )}	984	Prkd2	Serine/threonine-protein kinase D2	1.2605

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	985	Eif4ebp1	Eukaryotic translation initiation	1.2599
	GGTLFST*TPGGTR{circumflex over ( )}			factor 4E-binding protein 1

IPI:IPI00230368.1	SSSLGNSPDR{circumflex over ( )}GPLR{circumflex over ( )}PF	986	2610110G12Rik	Isoform 1 of UPF0635 protein C6orf134	1.2571
	VPEQELLR{circumflex over ( )}			homolog

IPI:IPI00845596.1	LFS*QGQDVSDK@VK@	987	Mllt4	Isoform	3 of Afadin	1.2551

IPI:IPI00378026.4	VSSPSCDSQGLHPEEAPS	988	Nacc1	Nucleus accumbens-associated protein 1	1.253
	SEPQSPVAQTLGWPACSTP
	LPLVSR{circumflex over ( )}

IPI:IPI00458460.1	SLSESSVVMDR{circumflex over ( )}	989	Zfp106	Isoform	1 of Zinc finger protein 106	1.2522

IPI:IPI00399440.2	HLEDAGSTPSIGENDLK@	990	Rictor	Isoform	1 of Rapamycin-insensitive	1.2463
	FPK@			companion of mTOR

IPI:IPI00109318.1	TVAISDAAQLPQDYCTT*PG	991	Eif4ebp2	Eukaryotic translation initiation	1.2423
	GTLFSTT*PGGTR{circumflex over ( )}IIYDR{circumflex over ( )}K@			factor 4E-binding protein 2

IPI:IPI00109318.1	TVAISDAAQLPQDYCTT*PG	992	Eif4ebp2	Eukaryotic translation initiation	1.2423
	GTLFST*TPGGTRIIYDRK			factor 4E-binding protein 2

IPI:IPI00379682.1	R{circumflex over ( )}ANTLS*HFPVECPAPPEP	993	Tbc1d1	Isoform	1 of TBC1 domain family	1.2355
	AQS*SPGVSQR{circumflex over ( )}			member 1

IPI:IPI00454104.1	TTEAPCSPGSQQPPSPDE	994	Scrib	Isoform	1 of Protein LAP4	1.2352
	LPANVK

IPI:IPI00454104.1	TTEAPCSPGSQQPPSPDE	995	Scrib	Isoform	1 of Protein LAP4	1.2352
	LPANVK@

IPI:IPI00454104.1	TTEAPCSPGSQQPPSPDE	996	Scrib	Isoform 1 of Protein LAP4	1.2352
	LPANVK@

IPI:IPI00109318.1	TVAISDAAQLPQDYCT*TPG	997	Eif4ebp2	Eukaryotic translation initiation	1.2336
	GTLFSTT*PGGTR{circumflex over ( )}IIYDR{circumflex over ( )}K@			factor 4E-binding protein 2

IPI:IPI00378438.6	S*QSVPGAWPGASPLSSQP	998	Tns1	tensin	1	1.2303
	LLGSS*R{circumflex over ( )}QSHPLTQSR{circumflex over ( )}

IPI:IPI00118143.1	KQTPPASPS*PQPIEDRPP	999	Cttn	Src substrate cortactin	1.2254
	SS*PIYEDAAPFKAEPSYR

IPI:IPI00330773.3	ASYSGTSPS*HSFISGEPDR{circumflex over ( )}	1000	Phldb2	Isoform	1 of Pleckstrin homology-like	1.2251
				domain family B member 2

IPI:IPI00330773.3	ASYSGTS*PSHSFISGEPDR{circumflex over ( )}	1001	Phldb2	Isoform	1 of Pleckstrin homology-like	1.2251
				domain family B member 2

IPI:IPI00127071.3	GATEEEQQDSGSEPR{circumflex over ( )}G	1002	Ddx41	Probable ATP-dependent RNA helicase	1.223
	DEDDIPLGPQSNVSLLDQH			DDX41
	QHLK@

IPI:IPI00153986.2	GTSR{circumflex over ( )}PGTPSAEAASTSS	1003	Gtf2f1	General transcription factor IIF	1.2226
	TLR{circumflex over ( )}			subunit 1

IPI:IPI00153986.2	GTSR{circumflex over ( )}PGTPSAEAASTSS	1004	Gtf2f1	General transcription factor IIF	1.2226
	TLR{circumflex over ( )}			subunit 1

IPI:IPI00153986.2	GTSRPGTPSAEAASTSST	1005	Gtf2f1	General transcription factor IIF	1.2226
	LR			subunit	1

IPI:IPI00117277.2	R{circumflex over ( )}GS*GDTSISMDTEASIR{circumflex over ( )}	1006	Lrrfip1	Isoform	2 of Leucine-rich repeat	1.2213
				flightless-interacting protein 1

IPI:IPI00661508.3	LYSLLGIDLTAPSNNS*SPR{circumflex over ( )}	1007	Sbno1	Isoform	2 of Protein strawberry notch	1.2147
	DS*PCK@ENK@			homolog 1

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	1008	Eif4ebp1	Eukaryotic translation initiation	1.2095
	*SPTSDEPPMQASQSQLPS			factor 4E-binding protein 1
	S*PEDK

IPI:IPI00116442.1	TRLASESANDDNEDS	1009	Hdgfrp2	Isoform	3 of Hepatoma derived growth	1.1963
				factor-related protein 2

IPI:IPI00116442.1	TRLASESANDDNEDS	1010	Hdgfrp2	Isoform	3 of Hepatoma-derived growth	1.1963
				factor-related protein 2

IPI:IPI00929761.1	R{circumflex over ( )}CPS*QSSSR{circumflex over ( )}PATGISQP	1011	Larp1	Isoform 2 of La-related protein 1	1.1959
	PTTPTGQATR{circumflex over ( )}

IPI:IPI00320905.7	LGEQGPEPGPTPPQTPT*	1012	Arhgap17	Isoform 1 of Rho GTPase-activating	1.1918
	PPS*TPPLAK@			protein 17

IPI:IPI00420509.2	S*IEDLQPPNALSAPFTNS	1013	Fmnl3	Isoform	1 of Formin-like protein 3	1.1892
	LAR

IPI:IPI00153986.2	GTSRPGTPSAEAASTSST	1014	Gtf2f1	General transcription factor IIF	1.1888
				subunit 1

IPI:IPI00153986.2	GTSR{circumflex over ( )}PGTPSAEAASTSS	1015	Gtf2f1	General transcription factor IIF	1.1888
	TLR{circumflex over ( )}			subunit 1

IPI:IPI00153986.2	GTSRPGTPSAEAASTSST	1016	Gtf2f1	General transcription factor IIF	1.1888
	LR			subunit	1

IPI:IPI00153986.2	GTSR{circumflex over ( )}PGTPSAEAASTSS	1017	Gtf2f1	General transcription factor IIF	1.1888
	TLR{circumflex over ( )}			subunit 1

IPI:IPI00320905.7	LGEQGPEPGPTPPQTPT*	1018	Arhgap17	Isoform 1 of Rho GTPase-activating	1.1882
	PPST*PPLAK			protein	17

IPI:IPI00330773.3	ASYSGTS*PSHSFISGEPDR	1019	Phldb2	Isoform	1 of Pleckstrin homology-like	1.1879
				domain family B member 2

IPI:IPI00330773.3	ASYSGTSPS*HSFISGEPDR{circumflex over ( )}	1020	Phldb2	Isoform	1 of Pleckstrin homology-like	1.1879
				domain family B member 2

IPI:IPI00330773.3	ASYSGTS*PSHSFISGEPDR{circumflex over ( )}	1021	Phldb2	Isoform	1 of Pleckstrin homology-like	1.1879
				domain family B member 2

IPI:IPI00330773.3	SDELLGDLTRT*PPSSSAAF	1022	Phldb2	Isoform	1 of Pleckstrin homology-like	1.185
	LK			domain family B member 2

IPI:IPI00330773.3	SDELLGDLTR{circumflex over ( )}T*PPSSSAA	1023	Phldb2	Isoform	1 of Pleckstrin homology-like	1.185
	FLK@			domain family B member 2

IPI:IPI00340860.5	TASEGSEAETPEAPK@Q	1024	Larp7	Isoform	1 of La-related protein 7	1.1845
	PAK@K@

IPI:IPI00340860.5	TASEGSEAETPEAPK@Q	1025	Larp7	Isoform	1 of La-related protein 7	1.1845
	PAK@K@

IPI:IPI00129264.1	ATSRPINLGPSS*PNTEIHW	1026	Sorbs3	Vinexin	1.175
	TPYR

IPI:IPI00129264.1	ATSR{circumflex over ( )}PINLGPSS*PNTEIHW	1027	Sorbs3	Vinexin	1.175
	TPYR{circumflex over ( )}

IPI:IPI00129264.1	ATSRPINLGPS*SPNTEIHW	1028	Sorbs3	Vinexin	1.175
	TPYR

IPI:IPI00381495.5	TVSSPIPYTPSPSSS*R{circumflex over ( )}PI	1029	Ccdc6	coiled-coil domain containing 6	1.1749
	SPGLSYASHTVGFT*PPTSL
	TR{circumflex over ( )}

IPI:IPI00153986.2	GTSR{circumflex over ( )}PGTPSAEAASTSS	1030	Gtf2f1	General transcription factor IIF	1.1748
	TLR{circumflex over ( )}			subunit 1

IPI:IPI00153986.2	GTSRPGTPSAEAASTSST	1031	Gtf2f1	General transcription factor IIF	1.1748
	LR			subunit	1

IPI:IPI00153986.2	GTSR{circumflex over ( )}PGTPSAEAASTSS	1032	Gtf2f1	General transcription factor IIF	1.1748
	TLR{circumflex over ( )}			subunit 1

IPI:IPI00153986.2	GTSRPGTPSAEAASTSST	1033	Gtf2f1	General transcription factor IIF	1.1748
	LR			subunit	1

IPI:IPI00136572.4	YEENPAWPGTSTHNGPNG	1034	Zc3hav1	Isoform 1 of Zinc finger CCCH-type	1.1738
	FSQIMDETPNVSK@SS*PT			antiviral protein 1
	GFGIK@

IPI:IPI00225062.2	GSLSR{circumflex over ( )}SSS*PVTELTAR{circumflex over ( )}	1035	Srrm2	Isoform	3 of Serine/arginine repetitive	1.1701
				matrix protein 2

IPI:IPI00133685.1	S*LPVSVPVWAFK@	1036	Akt1s1	Proline-rich AKT1 substrate 1	1.1669

IPI:IPI00133685.1	S*LPVSVPVWAFK@	1037	Akt1s1	Proline-rich AKT1 substrate 1	1.1669

IPI:IPI00133685.1	S*LPVSVPVWAFK@	1038	Akt1s1	Proline-rich AKT1 substrate 1	1.1669

IPI:IPI00225062.2	GSLSRSSS*PVTELTAR	1039	Srrm2	Isoform	3 of Serine/arginine repetitive	1.1669
				matrix protein 2

IPI:IPI00225062.2	GSLSR{circumflex over ( )}SSS*PVTELTAR{circumflex over ( )}	1040	Srrm2	Isoform	3 of Serine/arginine repetitive	1.1669
				matrix protein 2

IPI:IPI00929786.1	S*LPTTVPESPNYR{circumflex over ( )}	1041	Larp1	Isoform	1 of La-related protein 1	1.1655

IPI:IPI00929786.1	S*LPTTVPESPNYR	1042	Larp1	Isoform	1 of La-related protein 1	1.1655

IPI:IPI00116442.1	TRLASESANDDNEDS	1043	Hdgfrp2	Isoform	3 of Hepatoma-derived growth	1.1626
				factor-related protein 2

IPI:IPI00116442.1	TR{circumflex over ( )}LASESANDDNEDS	1044	Hdgfrp2	Isoform	3 of Hepatoma-derived growth	1.1626
				factor-related protein 2

IPI:IPI00116442.1	TR{circumflex over ( )}LASESANDDNEDS	1045	Hdgfrp2	Isoform	3 of Hepatoma-derived growth	1.1626
				factor-related protein 2

IPI:IPI00116442.1	TRLASESANDDNEDS	1046	Hdgfrp2	Isoform	3 of Hepatoma-derived growth	1.1626
				factor-related protein 2

IPI:IPI00317599.3	SQEDEEEIST*SPGVSEFVS	1047	Syap1	Synapse-associated protein 1	1.1567
	DAFDTCSLNQEDLRK

IPI:IPI00317599.3	SQEDEEEISTS*PGVSEFVS	1048	Syap1	Synapse-associated protein 1	1.1567
	DAFDTCSLNQEDLR{circumflex over ( )}K@

IPI:IPI00317599.3	SQEDEEEISTS*PGVSEFVS	1049	Syap1	Synapse-associated protein 1	1.1567
	DAFDTCSLNQEDLR{circumflex over ( )}K@

IPI:IPI00317599.3	SQEDEEEISTS*PGVSEFVS	1050	Syap1	Synapse-associated protein 1	1.1567
	DAFDTCSLNQEDLRK

IPI:IPI00126124.2	TFSECSYPETEEEAEALP	1051	St5	Isoform	1 of Suppression of	1.1565
	GR{circumflex over ( )}			tumorigenicity 5

IPI:IPI00312600.2	LSSDENSNPDLS*GDENDD	1052	Eed	Isoform	1 of Polycomb protein EED	1.1553
	AVSIESGTNTER{circumflex over ( )}PDTPTNT
	PNAPGR{circumflex over ( )}K@

IPI:IPI00312600.2	LSSDENSNPDLS*GDENDD	1053	Eed	Isoform 1 of Polycomb protein EED	1.1553
	AVSIESGTNTERPDTPTNTP
	NAPGRK

IPI:IPI00229990.7	RPPGDVLETFNFLENADDS*	1054	Strn3	Striatin-3	1.1544
	DEEENDMIEGIPEGKDK

IPI:IPI00229990.7	R{circumflex over ( )}PPGDVLETFNFLENADDS	1055	Strn3	Striatin-3	1.1544
	*DEEENDMIEGIPEGK@DK@

IPI:IPI00121418.1	DGEGPDNLEPACPLSLPLQ	1056	Rb1	Retinoblastoma-associated protein	1.152
	GNHTAADMYLSPLRSPK

IPI:IPI00121418.1	DGEGPDNLEPACPLSLPLQ	1057	Rb1	Retinoblastoma-associated protein	1.152
	GNHTAADMYLSPLR{circumflex over ( )}SPK@

IPI:IPI00121418.1	DGEGPDNLEPACPLSLPLQ	1058	Rb1	Retinoblastoma-associated protein	1.152
	GNHTAADMYLSPLR{circumflex over ( )}SPK@

IPI:IPI00136386.2	GDTSETASATPAY*R{circumflex over ( )}	1059	Tssk4	Isoform	2 of Testis-specific	1.15
				serine/threonine-protein kinase 4

IPI:IPI00515576.5	TGVNENTVVSAGKDLSTS*P	1060	Ehbp1	Isoform 2 of EH domain-binding	1.1492
	KPSPIPSPVLGQKPNASQ			protein 1
	SLLAWCR

IPI:IPI00420143.1	STSSVDSDILSSSHSSDTL	1061	Usp47	Isoform	2 of Ubiquitin carboxyl-	1.1474
	CNADSAQIPLANGLDSHSIT			terminal hydrolase 47
	SSR{circumflex over ( )}

IPI:IPI00109318.1	TVAISDAAQLPQDYCTT*PG	1062	Eif4ebp2	Eukaryotic translation initiation	1.147
	GTLFSTT*PGGTRIIYDRK			factor 4E-binding protein 2

IPI:IPI00109318.1	TVAISDAAQLPQDYCT*TPG	1063	Eif4ebp2	Eukaryotic translation initiation	1.147
	GTLFSTT*PGGTR{circumflex over ( )}IIYDR{circumflex over ( )}K@			factor 4E-binding protein 2

IPI:IPI00330773.3	SDELLGDLTR{circumflex over ( )}T*PPSSSAA	1064	Phldb2	Isoform	1 of Pleckstrin homology-like	1.1466
	FLK@			domain family B member 2

IPI:IPI00330773.3	SDELLGDLTRT*PPSSSAAF	1065	Phldb2	Isoform	1 of Pleckstrin homology-like	1.1466
	LK			domain family B member 2

IPI:IPI00229697.3	GTFSDQELDAQSLDDEDD	1066	—	68 kDa protein	1.1465
	SLQHAVHPALNRFSPSPR

IPI:IPI00133685.1	S*LPVSVPVWAFK@	1067	Akt1s1	Proline-rich AKT1 substrate 1	1.1417

IPI:IPI00133685.1	S*LPVSVPVWAFK	1068	Akt1s1	Proline-rich AKT1 substrate 1	1.1417

IPI:IPI00133685.1	S*LPVSVPVWAFK@	1069	Akt1s1	Proline-rich AKT1 substrate 1	1.1417

IPI:IPI00133685.1	S*LPVSVPVWAFK	1070	Akt1s1	Proline-rich AKT1 substrate 1	1.1417

IPI:IPI00130920.1	SLMSS*PEDLTK@DFEELK	1071	Mtap1b	Microtubule-associated protein 1B	1.1401
	@AEEIDVAK@

IPI:IPI00130920.1	SLMSSPEDLT*KDFEELKAE	1072	Mtap1b	Microtubule-associated protein 1B	1.1401
	EIDVAK

IPI:IPI00828355.1	GTLVHTTSDSDSEDGDQE	1073	Fmn1	Isoform	5 of Formin-1	1.1401
	AEEESSLDTQK@PTTVVLC
	EPSQEPK@

IPI:IPI00381495.5	TVSSPIPYTPSPSSSR{circumflex over ( )}PIS	1074	Ccdc6	coiled-coil domain containing 6	1.1376
	PGLSYASHTVGFTPPTSL
	TR{circumflex over ( )}

IPI:IPI00929761.1	CPS*QSSSRPATGISQPPTT	1075	Larp1	Isoform	2 of La-related protein 1	1.1372
	PTGQATR

IPI:IPI00379682.1	R{circumflex over ( )}ANTLS*HFPVECPAPPEP	1076	Tbc1d1	Isoform	1 of TBC1 domain family	1.1351
	AQSS*PGVSQR{circumflex over ( )}			member 1

IPI:IPI00379682.1	RANT*LSHFPVECPAPPEPA	1077	Tbc1d1	Isoform	1 of TBC1 domain family	1.1351
	QSS*PGVSQR			member	1

IPI:IPI00381495.5	TVSSPIPYTPSPSSSR{circumflex over ( )}PIS	1078	Ccdc6	coiled-coil domain containing 6	1.1342
	PGLSYASHTVGFTPPTSL
	TR{circumflex over ( )}

IPI:IPI00381495.5	TVSSPIPYTPSPSSSRPIS	1079	Ccdc6	coiled-coil domain containing 6	1.1342
	PGLSYASHTVGFTPPTSL
	TR

IPI:IPI00225062.2	M#SCFSR{circumflex over ( )}PSMSPTPLDR{circumflex over ( )}	1080	Srrm2	Isoform	3 of Serine/arginine repetitive	1.1322
				matrix protein 2

IPI:IPI00133374.5	EVDPSTGELQSLQMPESE	1081	Sqstm1	Isoform	1 of Sequestosome-1	1.1315
	GPS*SLDPSQEGPTGLK@

IPI:IPI00133374.5	EVDPSTGELQSLQMPESE	1082	Sqstm1	Isoform	1 of Sequestosome-1	1.1315
	GPS*SLDPSQEGPTGLK@

IPI:IPI00133374.5	EVDPSTGELQSLQMPESE	1083	Sqstm1	Isoform	1 of Sequestosome-1	1.1315
	GPS*SLDPSQEGPTGLK@

IPI:IPI00133374.5	EVDPSTGELQSLQMPESE	1084	Sqstm1	Isoform	1 of Sequestosome-1	1.1315
	GPS*SLDPSQEGPTGLK

IPI:IPI00465879.2	LCSSSSSDTSPR{circumflex over ( )}	1085	Zc3hc1	Isoform	1 of Nuclear-interacting	1.1277
				partner of ALK

IPI:IPI00468802.5	YEMACAIADAFNLPSSHLR{circumflex over ( )}	1086	Mat2b	Isoform	1 of Methionine	1.1262
	PITDS*PVIGAQR{circumflex over ( )}PK@			adenosyltransferase 2 subunit beta

IPI:IPI00121251.7	LSTTPSPTNSLHEDGVDD	1087	Tox4	TOX high mobility group box family	1.1209
	FR{circumflex over ( )}R{circumflex over ( )}			member 4

IPI:IPI00121251.7	LSTTPSPTNSLHEDGVDD	1088	Tox4	TOX high mobility group box family	1.1209
	FR{circumflex over ( )}R{circumflex over ( )}			member 4

IPI:IPI00121251.7	LSTTPSPTNSLHEDGVDD	1089	Tox4	TOX high mobility group box family	1.1209
	FRR			member	4

IPI:IPI00127554.1	R{circumflex over ( )}FSVQEQDWETTPPK@	1090	Emg1	Probable ribosome biogenesis protein	1.1179
	K@			NEP1

IPI:IPI00127554.1	RFSVQEQDWETTPPKK	1091	Emg1	Probable ribosome biogenesis protein	1.1179
				NEP1

IPI:IPI00459443.5	GEGVSQVGPGTPPAPES*P	1092	Tnks1bp1	182 kDa tankyrase-1-binding protein	1.1175
	RKPISGVQGNDPGISLPQR

IPI:IPI00225062.2	MELGTPLR{circumflex over ( )}HSGSTSPYP	1093	Srrm2	Isoform	3 of Serine/arginine repetitive	1.1138
	K@			matrix protein	2

IPI:IPI00225062.2	MELGTPLRHSGSTSPYPK	1094	Srrm2	Isoform	3 of Serine/arginine repetitive	1.1138
				matrix protein 2

IPI:IPI00225062.2	MELGTPLR{circumflex over ( )}HSGSTSPYP	1095	Srrm2	Isoform	3 of Serine/arginine repetitive	1.1138
	K@			matrix protein	2

IPI:IPI00127764.2	EISQSR{circumflex over ( )}NPSVSEHLPDEK@	1096	Pcm1	Isoform	1 of Pericentriolar material 1	1.1126
				protein

IPI:IPI00318938.6	NSPVAK@TPPK@DLPAIP	1097	Eif4ebp1	Eukaryotic translation initiation	1.1074
	GVTSPTSDEPPMQASQSQL			factor 4E-binding protein 1
	PSS*PEDK@

IPI:IPI00116923.2	SGFAR{circumflex over ( )}PGDLEFEDFS*QVI	1098	Trip10	Isoform 3 of Cdc42-interacting	1.1069
	NR{circumflex over ( )}VPSDSSLGT*PDGR{circumflex over ( )}PE			protein	4
	LR{circumflex over ( )}

IPI:IPI00121335.1	CGS*PSDSSTSEMMEVAVN	1099	LOC100048123;	RAC-beta serine/threonine-protein	1.1028
	K@		Akt2	kinase

IPI:IPI00356608.4	VGSLTPPS*SPK@	1100	Aak1	Isoform 2 of AP2-associated protein	1.1012
				kinase 1

IPI:IPI00828461.1	GPPDFSSDEER{circumflex over ( )}EPTPVL	1101	Tmpo	thymopoietin isoform delta	1.1001
	GSGASVGR{circumflex over ( )}

IPI:IPI00381495.5	TVSSPIPYTPSPSSSR{circumflex over ( )}PIS	1102	Ccdc6	coiled-coil domain containing 6	1.0993
	PGLSYASHTVGFTPPTSLTR{circumflex over ( )}

IPI:IPI00381495.5	TVSSPIPYTPS*PSSSRPISP	1103	Ccdc6	coiled-coil domain containing 6	1.0993
	GLSY*ASHTVGFTPPTSLTR

IPI:IPI00127554.1	R{circumflex over ( )}FS*VQEQDWETTPPK@K@	1104	Emg1	Probable ribosome biogenesis protein	1.0981
				NEP1

IPI:IPI00127554.1	R{circumflex over ( )}FS*VQEQDWETTPPK@K@	1105	Emg1	Probable ribosome biogenesis protein	1.0981
				NEP1

IPI:IPI00121251.7	LSTTPSPTNSLHEDGVDD	1106	Tox4	TOX high mobility group box family	1.0844
	FRR			member	4

IPI:IPI00121251.7	LSTTPSPTNSLHEDGVDD	1107	Tox4	TOX high mobility group box family	1.0844
	FR{circumflex over ( )}R{circumflex over ( )}			member 4

IPI:IPI00121251.7	LSTTPSPTNSLHEDGVDD	1108	Tox4	TOX high mobility group box family	1.0844
	FR{circumflex over ( )}R{circumflex over ( )}			member 4

IPI:IPI00318938.6	NSPVAK@TPPK@DLPAIP	1109	Eif4ebp1	Eukaryotic translation initiation	1.0842
	GVTSPTSDEPPMQASQSQL			factor 4E-binding protein 1
	PSS*PEDK@

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	1110	Eif4ebp1	Eukaryotic translation initiation	1.0842
	S*PTSDEPPMQASQSQLPS			factor 4E-binding protein 1
	SPEDK

IPI:IPI00169771.1	SSASGTHSESPEK@LQC	1111	BC031781	UPF0667 protein C1orf55 homolog	1.0823
	PVTEPGQGILENTGTEPGE
	TSDK@ECNER{circumflex over ( )}

IPI:IPI00318938.6	NSPVAK@TPPK@DLPAIP	1112	Eif4ebp1	Eukaryotic translation initiation	1.0778
	GVTSPTSDEPPMQASQSQL			factor 4E-binding protein 1
	PSS*PEDK@

IPI:IPI00127764.2	EISQSRNPSVSEHLPDEK	1113	Pcm1	Isoform	1 of Pericentriolar material 1	1.074
				protein

IPI:IPI00112597.1	SQSSEGVSSLSSSPSNSL	1114			1.0712
	ETQSQSLSR{circumflex over ( )}

IPI:IPI00112597.1	SQSSEGVSSLSSSPSNSL	1115			1.0712
	ETQSQSLSR{circumflex over ( )}

IPI:IPI00225062.2	SRSSSPDSKMELGTPLR	1116	Srrm2	Isoform	3 of Serine/arginine repetitive	1.0696
				matrix protein 2

IPI:IPI00330773.3	ASYSGTSPSHSFISGEPD	1117	Phldb2	Isoform 1 of Pleckstrin homology-like	1.0688
	R{circumflex over ( )}			domain family B member 2

IPI:IPI00330246.2	WAAHGTS*PEDFSLTLGAR{circumflex over ( )}	1118	Phldb1	Isoform	2 of Pleckstrin homology-like	1.068
				domain family B member 1

IPI:IPI00122594.4	NGVSLFNSPK@T*EQPSPV	1119	Ahctf1	AT-hook-containing transcription	1.0661
	VHSFPHPELPEAFVGTPISN			factor 1
	TSQR{circumflex over ( )}

IPI:IPI00122594.4	NGVSLFNS*PKTEQPSPVV	1120	Ahctf1	AT-hook-containing transcription	1.0661
	HSFPHPELPEAFVGTPISNT			factor 1
	SQR

IPI:IPI00122594.4	NGVSLFNSPK@TEQPSPVV	1121	Ahctf1	AT-hook-containing transcription	1.0661
	HS*FPHPELPEAFVGTPISN			factor 1
	TSQR{circumflex over ( )}

IPI:IPI00121418.1	DGEGPDNLEPACPLSLPLQ	1122	Rb1	Retinoblastoma-associated protein	1.0618
	GNHTAADMYLSPLR{circumflex over ( )}SPK@

IPI:IPI00121418.1	DGEGPDNLEPACPLSLPLQ	1123	Rb1	Retinoblastoma-associated protein	1.0618
	GNHTAADMYLSPLRSPK

IPI:IPI00320905.7	LGEQGPEPGPTPPQTPT*	1124	Arhgap17	Isoform 1 of Rho GTPase-activating	1.0618
	PPST*PPLAK			protein 17

IPI:IPI00403116.9	NGS*PVPVPDISQEPDGPAL	1125	Ncaph2	Isoform 3 of Condensin-2 complex	1.0598
	S*GGEEDAEDGAEPLEVAL			subunit H2
	EPAEPR{circumflex over ( )}

IPI:IPI00403116.9	NGS*PVPVPDISQEPDGPAL	1126	Ncaph2	Isoform 3 of Condensin-2 complex	1.0598
	S*GGEEDAEDGAEPLEVAL			subunit H2
	EPAEPR

IPI:IPI00403116.9	NGSPVPVPDISQEPDGPA	1127	Ncaph2	Isoform 3 of Condensin-2 complex	1.0598
	LSGGEEDAEDGAEPLEVAL			subunit H2
	EPAEPR{circumflex over ( )}

IPI:IPI00230237.1	TAVTATNVSAHGSQANS*P	1128	Pbx1	Isoform PBX1b of Pre-B-cell leukemia	1.054
	STPNSAGGY*PSPCYQPDR			transcription factor 1
	{circumflex over ( )}R{circumflex over ( )}

IPI:IPI00421139.8	SS*SHLDPAATPHSTLQGSS	1129	Dlg5	Putative uncharacterized protein	1.0498
	AGTPEHPSVIDPLMEQDEG
	PGTPPAK@

IPI:IPI00121251.7	LSTTPSPTNSLHEDGVDD	1130	Tox4	TOX high mobility group box family	1.0342
	FRR			member	4

IPI:IPI00121251.7	LSTTPSPTNSLHEDGVDD	1131	Tox4	TOX high mobility group box family	1.0342
	FR{circumflex over ( )}R{circumflex over ( )}			member 4

IPI:IPI00121431.4	S*INYSELDQFPSELEK	1132	Hells	Isoform 1 of Lymphocyte specific	1.0332
				helicase

IPI:IPI00121431.4	S*INYSELDQFPSELEK@	1133	Hells	Isoform 1 of Lymphocyte-specific	1.0332
				helicase

IPI:IPI00121431.4	S*INYSELDQFPSELEK@	1134	Hells	Isoform 1 of Lymphocyte-specific	1.0332
				helicase

IPI:IPI00454039.3	VGS*EHSLLDPPGK	1135	Erbb2ip	Isoform 1 of Protein LAP2	1.0307

IPI:IPI00109318.1	TVAISDAAQLPQDYCTT*PG	1136	Eif4ebp2	Eukaryotic translation initiation	1.0271
	GTLFSTT*PGGTR{circumflex over ( )}IIYDR{circumflex over ( )}			factor 4E-binding protein 2

IPI:IPI00109318.1	TVAISDAAQLPQDYCTT*PG	1137	Eif4ebp2	Eukaryotic translation initiation	1.0271
	GTLFSTT*PGGTRIIYDR			factor 4E-binding protein 2

IPI:IPI00226228.5	ANNAGASIYPTGPADPCPP	1138	Azi1	5-azacytidine-induced protein 1	1.0268
	ASESSPEQWQSPEDKPQ
	DIHSQGEAR

IPI:IPI00317401.6	AESPETSAVESTQSTPQK@	1139	Pds5b	Isoform	1 of Sister chromatid cohesion	1.0246
				protein PDS5 homolog B

IPI:IPI00317401.6	AESPETSAVESTQSTPQK@	1140	Pds5b	Isoform	1 of Sister chromatid cohesion	1.0246
				protein PDS5 homolog B

IPI:IPI00317401.6	AESPETSAVESTQSTPQK	1141	Pds5b	Isoform	1 of Sister chromatid cohesion	1.0246
				protein PDS5 homolog B

IPI:IPI00113389.5	HNLFEDNM#ALPSESVSS*L	1142	Fam129a	Protein Niban	1.0232
	TDLK

IPI:IPI00318938.6	NSPVAK@TPPK@DLPAIP	1143	Eif4ebp1	Eukaryotic translation initiation	1.0227
	GVTSPTSDEPPM#QASQSQ			factor 4E-binding protein 1
	LPSSPEDK@

IPI:IPI00463074.6	SRSSPSTDHYSQEVPVEP	1144	Shroom2	shroom family member 2	1.0176
	NR

IPI:IPI00463074.6	SRSSPSTDHYSQEVPVEP	1145	Shroom2	shroom family member 2	1.0176
	NR

IPI:IPI00463074.6	SR{circumflex over ( )}SSPSTDHYSQEVPVE	1146	Shroom2	shroom family member 2	1.0176
	PNR{circumflex over ( )}

IPI:IPI00553798.2	SNSFSDERAEFSAPSTPT	1147	Ahnak	AHNAK nucleoprotein isoform 1	1.0143
	GTLEFAGGDAK@

IPI:IPI00553798.2	SNSFSDEREFSAPSTPTG	1148	Ahnak	AHNAK nucleoprotein isoform 1	1.0143
	TLEFAGGDAK

IPI:IPI00553798.2	SNSFSDEREFSAPSTPTG	1149	Ahnak	AHNAK nucleoprotein isoform 1	1.0143
	TLEFAGGDAK

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	1150	Eif4ebp1	Eukaryotic translation initiation	1.0129
	SPT*SDEPPMQASQSQLPS			factor 4E-binding protein 1
	S*PEDK

IPI:IPI00127131.6	GPSPSPVGSPASVAQSR{circumflex over ( )}	1151	Arid1a	AT rich interactive domain 1A	1.0107

IPI:IPI00113389.5	HNLFEDNMALPSES*VSSLT	1152	Fam129a	Protein Niban	1.0048
	DLK@TAMGSNQASPAR{circumflex over ( )}
	R{circumflex over ( )}

IPI:IPI00113389.5	HNLFEDNMALPSESVSSLT*	1153	Fam129a	Protein Niban	1.0048
	DLK@TAMGSNQASPAR{circumflex over ( )}
	R{circumflex over ( )}

IPI:IPI00454109.2	FPPSTPSEVLSPTEDPRS*	1154	Erf	ETS domain-containing transcription	1.0039
	PPACSSSSSSLFSAVVAR			factor ERF

IPI:IPI00120095.2	CDGSPR{circumflex over ( )}TPPS*TPPATAN	1155	Samhd1	SAM domain and HD domain-	1.0027
	LSADDDFQNTDLR{circumflex over ( )}			containing protein 1

IPI:IPI00117229.3	TS*PAGGTWSSVVSGVPR{circumflex over ( )}	1156	Atxn2	ataxin	2	1.0015

IPI:IPI00117229.3	TS*PAGGTWSSVVSGVPR	1157	Atxn2	ataxin	2	1.0015

IPI:IPI00320594.5	SQDSYPGSPSLSPR	1158	Ranbp10	Ran-binding protein 10	1.0009

IPI:IPI00320594.5	SQDSYPGSPSLSPR{circumflex over ( )}	1159	Ranbp10	Ran-binding protein 10	1.0009

IPI:IPI00320594.5	SQDSYPGSPSLSPR{circumflex over ( )}	1160	Ranbp10	Ran-binding protein 10	1.0009

IPI:IPI00320594.5	SQDSYPGSPSLSPR	1161	Ranbp10	Ran-binding protein 10	1.0009

IPI:IPI00320594.5	SQDSYPGSPSLSPR{circumflex over ( )}	1162	Ranbp10	Ran-binding protein 10	1.0009

IPI:IPI00121251.7	LSTTPSPTNSLHEDGVDD	1163	Tox4	TOX high mobility group box family	1.0005
	FR{circumflex over ( )}R{circumflex over ( )}			member 4

IPI:IPI00121251.7	LSTTPSPTNSLHEDGVDD	1164	Tox4	TOX high mobility group box family	1.0005
	FRR			member	4

Table 7. Phosphopeptides whose intensities decrease after rapamycin treatment (Rapamycin screen).

TABLE 8

					Log2
		SEQ			Area
		ID	Gene		(H/L)
Reference	Ascore Seq	NO	Name	Annotation	Ratio

Ku (light cells: control (rapamycin treated); heavy cells: treated with

rapamycin + Ku-0063794. L, light; H, heavy)

IPI:IPI00648918.3	DQEEELDEQAGS*IQM#LEQ	1165	Myo18a	Isoform 4 of Myosin-XVIIIa	−7.171
	LK

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1166	Eif4ebp1	Eukaryotic translation	−7.159
	GGTLFSTT*PGGTR			initiation factor 4E-binding protein 1

IPI:IPI00318938.6	VALGDGVQLPPGDYSTTP	1167	Eif4ebp1	Eukaryotic translation initiation	−7.127
	GGTLFSTTPGGTR			factor 4E-binding protein 1

IPI:IPI00622700.2	TAPPVLPTGYDS*EEEEESR	1168	Brd2	Isoform	2 of Bromodomain-containing	−6.823
	{circumflex over ( )}PMSYDEK@			protein 2

IPI:IPI00458153.2	ENPPSPPT*SPAAPQPR{circumflex over ( )}	1169	6330577E15Rik	Uncharacterized protein C10orf78	−6.819
				homolog

IPI:IPI00454090.1	DSVDLQS*LM#TEM#NR	1170	Best3	Bestrophin-3	−6.498

IPI:IPI00274140.4	LSPFFTLDLS*PTDDKSSKP	1171	Grit	Isoform 2 of Rho/Cdc42/Rac GTPase-	−6.465
	SSFTEK			activating protein RICS

IPI:IPI00929786.1	SLPTTVPESPNYR	1172	Larp1	Isoform	1 of La-related protein 1	−6.159

IPI:IPI00224969.2	VAPSMSSLNSLASSCFD	1173	Fam62c	Isoform 1 of Extended synaptotagmin-3	−6.023
	LTDVSLNTEAGDSR

IPI:IPI00929786.1	SLPTTVPESPNYR	1174	Larp1	Isoform 1 of La-related protein 1	−5.98

IPI:IPI00458039.3	IFTEANLVSVGSKK	1175	Mdn1	Midasin homolog	−5.907

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	1176	Eif4ebp1	Eukaryotic translation initiation	−5.874
	SPTSDEPPMQASQSQLPS*			factor 4E-binding protein 1
	SPEDK

IPI:IPI00130920.1	TLEVVSPSQSVTGS*AGHTP	1177	Mtap1b	Microtubule-associated protein 1B	−5.86
	YYQS*PTDEK

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1178	Eif4ebp1	Eukaryotic translation initiation	−5.685
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00270767.3	VVK@EDGVMS*PEK@	1179	Rtn4	Isoform	2 of Reticulon-4	−5.681

IPI:IPI00121430.2	DLKPETDYVVNVYSVVED	1180	Col12a1	Isoform	1 of Collagen alpha-1(XII)	−5.655
	EYSEPLKGT*EK			chain

IPI:IPI00169506.7	QAVYYLTLQAT*DGGNQS	1181	Pcdh24	Pcdh24 protein	−5.499
	TT*TALEITLLDINDNPPVVR

IPI:IPI00133685.1	LNT*SDFQK	1182	Akt1s1	Proline-rich AKT1 substrate 1	−5.419

IPI:IPI00876080.1	LPDLRS*R	1183	Gpr157	Putative uncharacterized protein	−5.334
				(Fragment)

IPI:IPI00850983.1	TPGPPS*SQGSPVDTQPAA	1184	Synj1	similar to mKIAA0910 protein	−5.321
	QK

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	1185	Eif4ebp1	Eukaryotic translation initiation	−4.991
	SPTSDEPPMQASQSQLPSS			factor 4E-binding protein 1
	*PEDK

IPI:IPI00282808.7	TPGVPKQKPCSPLSEPDA	1186	Zfp318	zinc finger protein 318 isoform 1	−4.923
	FLK

IPI:IPI00133374.5	EVDPSTGELQSLQMPESEG	1187	Sqstm1	Isoform	1 of Sequestosome-1	−4.888
	PS*SLDPSQEGPTGLK@

IPI:IPI00133374.5	EVDPSTGELQSLQMPESEG	1188	Sqstm1	Isoform	1 of Sequestosome-1	−4.882
	PS*SLDPSQEGPTGLK@

IPI:IPI00133685.1	LNT*SDFQK	1189	Akt1s1	Proline-rich AKT1 substrate 1	−4.871

IPI:IPI00623114.5	DMPAAGSLGSSSR	1190	Fat1	FAT tumor suppressor homolog 1	−4.85

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1191	Eif4ebp1	Eukaryotic translation initiation	−4.78
	GGT*LFSTTPGGTR			factor 4E-binding protein 1

IPI:IPI00125442.4	VPMVSSQPTSSM#VPPPI	1192	Alpk3	myocyte induction differentiation	−4.621
	KPLNR			originator

IPI:IPI00380641.3	KLPVVSS*VVK	1193	Zc3h14	Isoform	2 of Zinc finger CCCH domain-	−4.368
				containing protein 14

IPI:IPI00338745.4	QADVADQQTTELPAENGET	1194	Hmgn1;	Non-histone chromosomal protein	−4.324
	ENQSPAS*EEEKEAK		LOC100044391	HMG-14

IPI:IPI00338745.4	QADVADQQTTELPAENGET	1195	Hmgn1;	Non-histone chromosomal protein	−4.324
	ENQS*PASEEEKEAK		LOC100044391	HMG-14

IPI:IPI00271998.4	GPNS*PPPGMPLR	1196	2310057M21Rik	Uncharacterized protein C10orf88	−4.316
				homolog

IPI:IPI00338976.3	FTISAIS*K	1197	Fcrls	Fc receptor-like protein 2 scavenger	−4.254
				isoform

IPI:IPI00352984.4	IVHFSVNACLTPICSLHHV	1198	Xdh	Xanthine dehydrogenase/oxidase	−4.21
	AVTT*VEGIGNTKK

IPI:IPI00318938.6	VALGDGVQLPPGDYS*TTP	1199	Eif4ebp1	Eukaryotic translation initiation	−4.196
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00330246.2	K@GSFSGR{circumflex over ( )}LSPAYSLGS	1200	Phldb1	Isoform	2 of Pleckstrin homology-like	−4.16
	LTGASPR{circumflex over ( )}			domain family B member 1

IPI:IPI00330066.5	T*RSPDVISSASTALSQDIPE	1201	Edc4	Isoform	1 of Enhancer of mRNA-	−4.045
	IASEALSR			decapping protein 4

IPI:IPI00225062.2	RKETPS*PR	1202	Srrm2	Isoform 3 of Serine/arginine repetitive	−4.005
				matrix protein 2

IPI:IPI00405665.6	K@IIETM#SS*PK@	1203	Kif20b	Isoform 1 of M-phase phosphoprotein 1	−3.928

IPI:IPI00311187.4	SAASGSSGES*MDSVSVS	1204	Rgl1	Ral guanine nucleotide dissociation	−3.911
	SCESNHS*EAEEGPVTPM			stimulator-like 1
	DTPDEPQKK

IPI:IPI00153724.1	VHS*PPASLVPR{circumflex over ( )}	1205	1110007A13Rik	UPF0557 protein C10orf119 homolog	−3.905

IPI:IPI00121277.1	VAAAAGSGPSPPCSPGH	1206	Pi4k2a	Phosphatidylinositol 4-kinase type 2-	−3.904
	DR			alpha

IPI:IPI00331556.5	MQVDQEEPHTEEQQQQPQ	1207	Hspa4	Heat shock 70 kDa protein 4	−3.897
	TPAENKAESEEM#ET*SQA
	GSK

IPI:IPI00135443.2	KTSFDQDS*DVDIFPSDFTS	1208	Top2b	DNA topoisomerase 2-beta	−3.873
	EPPALPR

IPI:IPI00469938.3	VGSGAEPGEGS*PGSR{circumflex over ( )}PG	1209	BC021381	Isoform 2 of Uncharacterized protein	−3.843
	PIQADS*PK@			KIAA1931

IPI:IPI00331361.2	SPT*KAEPATPAEAAQSDR	1210	Mybbp1a	Myb-binding protein 1A	−3.83

IPI:IPI00625898.3	RQSSESDIESVMYT	1211	Bbx	Isoform	1 of HMG box transcription	−3.809
	IEAVAK			factor BBX

IPI:IPI00405227.3	S*LGEIAALTSK	1212	Vcl	Vinculin	−3.804

IPI:IPI00109318.1	RNS*PMAQTPPCHLPNIPGV	1213	Eif4ebp2	Eukaryotic translation initiation	−3.791
	TSPGALIEDSK			factor 4E-binding protein 2

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1214	Eif4ebp1	Eukaryotic translation initiation	−3.751
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	RVALGDGVQLPPGDY*STT	1215	Eif4ebp1	Eukaryotic translation initiation	−3.726
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	1216	Eif4ebp1	Eukaryotic translation initiation	−3.726
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	VALGDGVQLPPGDYSTTPG	1217	Eif4ebp1	Eukaryotic translation initiation	−3.705
	GTLFSTTPGGTR			factor 4E-binding protein 1

IPI:IPI00929814.1	KEANEVLSDSAGEDHPAE	1218	Morc2a	microrchidia 2A isoform 2	−3.673
	LR

IPI:IPI00229645.2	S*NDDLLAGMAGGVNVTNG	1219	Cytsa	Cytospin-A	−3.632
	IK

IPI:IPI00553798.2	AGAIS*ASGPELEGAGHSK	1220	Ahnak	AHNAK nucleoprotein isoform 1	−3.608

IPI:IPI00756424.3	TAR{circumflex over ( )}PNSEAPLSGSEDAD	1221	Eif5b	Isoform 1 of Eukaryotic translation	−3.543
	DSNK@			initiation factor 5B

IPI:IPI00121760.5	LKTEEGEIVY*SAEESENR	1222	Hnrpll	Isoform	1 of Heterogeneous nuclear	−3.51
				ribonucleoprotein L-like

IPI:IPI00458958.2	DWDKES*EGEEPAGGR	1223	Rrp15	RRP15-like protein	−3.46

IPI:IPI00458958.2	DWDK@ES*EGEEPAGGR{circumflex over ( )}	1224	Rrp15	RRP15-like protein	−3.46

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	1225	Eif4ebp1	Eukaryotic translation initiation	−3.429
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTTP	1226	Eif4ebp1	Eukaryotic translation initiation	−3.429
	GGTLFSTTPGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	1227	Eif4ebp1	Eukaryotic translation initiation	−3.429
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00109695.3	IEILDS*PASK	1228	D10Wsu102e	Uncharacterized protein C12orf45	−3.403
				homolog

IPI:IPI00230719.8	RDS*SDDWEIPDGQITVGQR	1229	Braf	Isoform 1 of B-Raf proto-oncogene	−3.372
				serine/threonine-protein kinase

IPI:IPI00377925.2	SS*PVCSTAPVETEPK	1230	Atg2b	Isoform 1 of Autophagy-related protein	−3.353
				2 homolog B

IPI:IPI00221723.1	MGQEFVES*K	1231	Wasf2	Wiskott-Aldrich syndrome protein family	−3.314
				member 2

IPI:IPI00165794.1	LWTHTSDQQVSAISNPSPC	1232	Tmem48	Nucleoporin NDC1	−3.255
	AS*VTAEGK

IPI:IPI00848747.1	DDSTCLGGILPAGLEPGS	1233	4930568B11Rik	hypothetical protein LOC75873	−3.253
	STR

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1234	Eif4ebp1	Eukaryotic translation initiation	−3.239
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	1235	Npm1	Nucleophosmin	−3.232
	AES*EDEDEEDVK@

IPI:IPI00318938.6	VALGDGVQLPPGDYST*TP	1236	Eif4ebp1	Eukaryotic translation initiation	−3.209
	GGTLFST*TPGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	VALGDGVQLPPGDYS*TTP	1237	Eif4ebp1	Eukaryotic translation initiation	−3.209
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1238	Eif4ebp1	Eukaryotic translation initiation	−3.175
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1239	Eif4ebp1	Eukaryotic translation initiation	−3.175
	GGTLFS*TTPGGTR			factor 4E-binding protein 1

IPI:IPI00120133.1	EDGDILASSFSVDTQVAYI	1240	Nat15	N-acetyltransferase 15	−3.163
	LSLGVVK

IPI:IPI00154109.2	SNSAPLIHGLSDSSPVFQA	1241	Fam122a	Protein FAM122A	−3.124
	EAPSAR

IPI:IPI00138199.5	DGYYGESTS*GR{circumflex over ( )}	1242	Pcsk5	proprotein convertase subtilisin/kexin	−3.103
				type 5

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	1243	Eif4ebp1	Eukaryotic translation initiation	−3.092
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00130920.1	TELS*PSFINPNPLEWFAGE	1244	Mtap1b	Microtubule-associated protein 1B	−3.089
	EPTEESEKPLTQSGGAPPP
	SGGK

IPI:IPI00131725.3	HQIVCINNDLSEEESDDES	1245	Npm3	Nucleoplasmin-3	−3.087
	*EEDEIK@LCGILPAK@

IPI:IPI00330066.5	T*RSPDVISSASTALSQDIPE	1246	Edc4	Isoform	1 of Enhancer of mRNA-	−3.084
	IASEALSR			decapping protein 4

IPI:IPI00756765.3	ALVIQESESPPSPPPSPDR	1247	Ehmt2	Euchromatic histone lysine N-	−3.071
	{circumflex over ( )}R{circumflex over ( )}			methyltransferase 2

IPI:IPI00221889.2	ILS*DDEDEDEEDAFK	1248	Ercc6l	DNA excision repair protein ERCC-6-	−3.063
				like

IPI:IPI00130920.1	TELS*PSFINPNPLEWFAGE	1249	Mtap1b	Microtubule-associated protein 1B	−3.052
	EPTEESEKPLTQSGGAPPP
	SGGK

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1250	Eif4ebp1	Eukaryotic translation initiation	−3.033
	GGTLFSTTPGGT*R			factor 4E-binding protein 1

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	1251	Eif4ebp1	Eukaryotic translation initiation	−3.031
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	1252	Eif4ebp1	Eukaryotic translation initiation	−3.031
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	1253	Eif4ebp1	Eukaryotic translation initiation	−3.031
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00137465.1	S*DGSLDDGDGIHR	1254	Serinc1	Serine incorporator 1	−3.027

IPI:IPI00130920.1	QGFPDRES*PVSDLTSTGLY	1255	Mtap1b	Microtubule-associated protein 1B	−3
	QDK

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1256	Eif4ebp1	Eukaryotic translation initiation	−2.995
	GGTLFSTTPGGT*R			factor 4E-binding protein 1

IPI:IPI00109318.1	RNSPMAQT*PPCHLPNIPGV	1257	Eif4ebp2	Eukaryotic translation initiation	−2.988
	TSPGALIEDSK			factor 4E-binding protein 2

IPI:IPI00122757.1	TDYYPEEMS*PPLMNPVSP	1258	Klf3;	Krueppel-like factor 3	−2.986
	PQALLQENHPS*VIVQPGK@		LOC100046855

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1259	Eif4ebp1	Eukaryotic translation initiation	−2.972
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00126006.6	TQT*PPLGQTPQLGLK	1260	Eif4g2	Isoform 1 of Eukaryotic translation	−2.956
				initiation factor 4 gamma 2

IPI:IPI00460668.2	KAENAEGQTPAIGPDGEPL	1261	Smarca4	Putative uncharacterized protein	−2.915
	DETSQM#S*DLPVK

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	1262	Eif4ebp1	Eukaryotic translation initiation	−2.908
	SPTSDEPPMQASQSQLPS*			factor 4E-binding protein 1
	SPEDK

IPI:IPI00127764.2	EGENVKPVNSGTWVAS*NS	1263	Pcm1	Isoform	1 of Pericentriolar material 1	−2.9
	ELTPSESLVTTDDETFEK			protein

IPI:IPI00874995.2	AS*DDLGEPDVFATAPFR	1264	Aak1	Uncharacterized protein FLJ45252	−2.899
				homolog

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	1265	Eif4ebp1	Eukaryotic translation initiation	−2.888
	SPTSDEPPMQASQSQLPSS			factor 4E-binding protein 1
	PEDK

IPI:IPI00318938.6	VALGDGVQLPPGDYSTTPG	1266	Eif4ebp1	Eukaryotic translation initiation	−2.884
	GTLFSTTPGGTR			factor 4E-binding protein 1

IPI:IPI00128564.1	R{circumflex over ( )}AAAASAAEAGIAT*PGTE	1267	Psmd4	Isoform Rpn10A of 26S proteasome	−2.861
	DSDDALLK@			non-ATPase regulatory subunit 4

IPI:IPI00318938.6	VALGDGVQLPPGDYSTTPG	1268	Eif4ebp1	Eukaryotic translation initiation	−2.846
	GTLFSTTPGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1269	Eif4ebp1	Eukaryotic translation initiation	−2.84
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00406741.5	AAVGVTGNDITTPPNKEPPP	1270	Mtap4	Isoform	4 of Microtubule-associated	−2.832
	S*PEKK			protein 4

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1271	Eif4ebp1	Eukaryotic translation initiation	−2.799
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1272	Eif4ebp1	Eukaryotic translation initiation	−2.799
	GGTLFST*TPGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1273	Eif4ebp1	Eukaryotic translation initiation	−2.794
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	VALGDGVQLPPGDYSTTPG	1274	Eif4ebp1	Eukaryotic translation initiation	−2.776
	GTLFSTTPGGTR			factor 4E-binding protein 1

IPI:IPI00109318.1	NS*PMAQTPPCHLPNIPGVT	1275	Eif4ebp2	Eukaryotic translation initiation	−2.763
	SPGALIEDSK			factor 4E-binding protein 2

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1276	Eif4ebp1	Eukaryotic translation initiation	−2.762
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1277	Eif4ebp1	Eukaryotic translation initiation	−2.756
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00331177.2	VSS*TPETPLTK@	1278	Pola2	DNA polymerase alpha subunit B	−2.747

IPI:IPI00109318.1	NSPMAQT*PPCHLPNIPGVT	1279	Eif4ebp2	Eukaryotic translation initiation	−2.714
	SPGALIEDSK			factor 4E-binding protein 2

IPI:IPI00318938.6	VALGDGVQLPPGDYSTTPG	1280	Eif4ebp1	Eukaryotic translation initiation	−2.672
	GTLFSTTPGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1281	Eif4ebp1	Eukaryotic translation initiation	−2.672
	GGTLFSTTPGGT*R			factor 4E-binding protein 1

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1282	Eif4ebp1	Eukaryotic translation initiation	−2.672
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00322707.5	IK@PVTENLVLPSHTGFCQ	1283	Atrx	Transcriptional regulator ATRX	−2.663
	SSGDEALSK@

IPI:IPI00322707.5	IK@PVTENLVLPSHTGFCQ	1284	Atrx	Transcriptional regulator ATRX	−2.663
	SSGDEALSK@

IPI:IPI00918973.1	IGFPETAEEELEEIASENS*D	1285	Strap	Serine-threonine kinase receptor-	−2.612
	SIYSSTPEVK			associated protein

IPI:IPI00318938.6	VALGDGVQLPPGDYS*TTP	1286	Eif4ebp1	Eukaryotic translation initiation	−2.565
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00125960.1	TASGSSVTS*LEGTR	1287	Ndrg1	Protein NDRG1	−2.557

IPI:IPI00125960.1	TASGSSVTS*LEGTR	1288	Ndrg1	Protein NDRG1	−2.557

IPI:IPI00125960.1	TASGSSVTSLEGTR	1289	Ndrg1	Protein NDRG1	−2.546

IPI:IPI00323045.3	S*MDVDLNQAHMEDTPK@	1290	Melk	Maternal embryonic leucine zipper	−2.53
				kinase

IPI:IPI00465761.5	SQLVHGCSQDSDELNPGGL	1291	Arhgef17	Isoform 1 of Rho guanine nucleotide	−2.526
	GSAGGVGSPEPPTSPR{circumflex over ( )}			exchange factor 17

IPI:IPI00465761.5	SQLVHGCSQDSDELNPGGL	1292	Arhgef17	Isoform	1 of Rho guanine nucleotide	−2.513
	GSAGGVGSPEPPTSPR{circumflex over ( )}			exchange factor 17

IPI:IPI00274140.4	LSPFFTLDLSPT*DDK@SSK	1293	Grit	Isoform	2 of Rho/Cdc42/Rac GTPase-	−2.507
	@PSSFTEK@			activating protein RICS

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	1294	Eif4ebp1	Eukaryotic translation initiation	−2.506
	PGGTLFSTFPGGTR			factor 4E-binding protein 1

IPI:IPI00314240.5	K@QPPVS*PGTALVGSQK@	1295	Hmga1	Isoform HMG-I of High mobility group	−2.493
				protein HMG-I/HMG-Y

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1296	Eif4ebp1	Eukaryotic translation initiation	−2.484
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00135379.3	VSPMLPSSSLES*PKDK	1297	Myo9b	Isoform 1 of Myosin-IXb	−2.452

IPI:IPI00753701.3	SQSGS*PAAPVEQVVIHTDT	1298	Mdc1	mediator of DNA damage checkpoint 1	−2.44
	SGDPTLPQR{circumflex over ( )}

IPI:IPI00221889.2	R{circumflex over ( )}ILS*DDEDEDEEDAFK@	1299	Ercc6l	DNA excision repair protein ERCC-6-	−2.438
				like

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	1300	Eif4ebp1	Eukaryotic translation initiation	−2.437
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	RVALGDGVQLPPGDYS*TT	1301	Eif4ebp1	Eukaryotic translation initiation	−2.437
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00454138.4	K@QQHVISTEEGDM#M#ET	1302	Sap130	Isoform 1 of Histone deacetylase	−2.434
	NSTDDEK@			complex subunit SAP130

IPI:IPI00454138.4	K@QQHVISTEEGDM#M#ET	1303	Sap130	Isoform 1 of Histone deacetylase	−2.434
	NSTDDEK@			complex subunit SAP130

IPI:IPI00753701.3	SQSGS*PAAPVEQVVIHTDT	1304	Mdc1	mediator of DNA damage checkpoint 1	−2.433
	SGDPTLPQR{circumflex over ( )}

IPI:IPI00654388.2	AENQR{circumflex over ( )}PAEDSALS*PGPLA	1305	Lrrfip1	Isoform	1 of Leucine-rich repeat	−2.425
	GAK@			flightless-interacting protein 1

IPI:IPI00654388.2	AENQRPAEDSALS*PGPLA	1306	Lrrfip1	Isoform	1 of Leucine-rich repeat	−2.425
	GAK			flightless-interacting protein 1

IPI:IPI00330121.4	GLPYADHNYGAPPPPT*PP	1307	Setd5	Isoform	2 of SET domain-containing	−2.414
	AS*PPVQTIIPR{circumflex over ( )}			protein 5

IPI:IPI00323256.3	HGS*SSYPPVIYSPLMPK	1308	Zfpm2	Isoform 1 of Zinc finger protein ZFPM2	−2.405

IPI:IPI00323256.3	HGSSS*YPPVIYSPLMPK	1309	Zfpm2	Isoform 1 of Zinc finger protein ZFPM2	−2.405

IPI:IPI00323256.3	HGSS*SYPPVIYSPLMPK@	1310	Zfpm2	Isoform	1 of Zinc finger protein ZFPM2	−2.395

IPI:IPI00123474.2	AGADIHAENEEPLCPLPSPS	1311	Nfkb2	NF-kB2 splice variant 4	−2.389
	TSGSDS*DSEGPER{circumflex over ( )}

IPI:IPI00117689.1	EGDELGEGERPEDDTAAIE	1312	Ptrf	Polymerase I and transcript release	−2.385
	LSSDEAVEVEEVIEESR			factor

IPI:IPI00117689.1	EGDELGEGERPEDDTAAIE	1313	Ptrf	Polymerase I and transcript release	−2.385
	LSSDEAVEVEEVIEESR			factor

IPI:IPI00127764.2	RSS*LVDEAPEDEEFEQK	1314	Pcm1	Isoform	1 of Pericentriolar material 1	−2.38
				protein

IPI:IPI00330121.4	GLPYADHNYGAPPPPT*PP	1315	Setd5	Isoform 2 of SET domain-containing	−2.359
	AS*PPVQTIIPR			protein 5

IPI:IPI00121277.1	VAAAAGSGPSPPCSPGH	1316	Pi4k2a	Phosphatidylinositol 4-kinase type 2-	−2.354
	DR			alpha

IPI:IPI00331361.2	SPT*KAEPATPAEAAQSDR	1317	Mybbp1a	Myb-binding protein 1A	−2.35

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	1318	Eif4ebp1	Eukaryotic translation initiation	−2.341
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00330121.4	GLPYADHNYGAPPPPT*PP	1319	Setd5	Isoform 2 of SET domain-containing	−2.334
	AS*PPVQTIIPIR{circumflex over ( )}			protein 5

IPI:IPI00623284.4	IWDPTPSHTPAGAATPGR	1320	Sf3b1	Splicing factor 3B subunit 1	−2.329
	GDTPGHATPGHGGATSS
	AR

IPI:IPI00153715.5	TEDEEFLIQHLLQAPS*PPR	1321	Prr12	proline rich 12	−2.321

IPI:IPI00116268.3	SGSCSSPQPKPNYPPLS	1322	N4bp1	N4bp1 protein	−2.307
	PPLPLPQLLPSVT*EAR

IPI:IPI00322707.5	AK@QPVIGDQNS*DSDEM#	1323	Atrx	Transcriptional regulator ATRX	−2.262
	LAVLK@

IPI:IPI00322707.5	AKQPVIGDQNS*DSDEM#LA	1324	Atrx	Transcriptional regulator ATRX	−2.262
	VLK

IPI:IPI00461244.3	DTTS*FEDISPQGISDDSST	1325	Ccdc88a	Isoform	2 of Girdin	−2.242
	GSR

IPI:IPI00317794.5	KEDSDEDEDEEDEDDSD	1326	Ncl	Nucleolin	−2.198
	EDEDDEEEDEFEPPIVK

IPI:IPI00317794.5	K@EDSDEDEDEEDEDDS	1327	Ncl	Nucleolin	−2.198
	DEDEDDEEEDEFEPPIVK@

IPI:IPI00400381.9	AHS*LLFENSDSFSEDTGTL	1328	Phactr4	Isoform	1 of Phosphatase and actin	−2.193
	GR{circumflex over ( )}			regulator 4

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAKD	1329	Eef1b2	Elongation factor 1-beta	−2.192
	DDDIDLFGS*DDEEESEEAK

IPI:IPI00923656.1	R{circumflex over ( )}DS*SSLSSPLNPK@	1330	Baz1b	Isoform 1 of Tyrosine-protein kinase	−2.17
				BAZ1B

IPI:IPI00318938.6	VALGDGVQLPPGDYSTTPG	1331	Eif4ebp1	Eukaryotic translation initiation	−2.17
	GTLFSTTPGGTR			factor 4E-binding protein 1

IPI:IPI00108389.5	ETVVSGPLGVEDISPSMS*P	1332	Trp53bp1	Transformation related protein 53	−2.161
	DDK@			binding protein 1

IPI:IPI00415558.1	AASPAKPS*SLDLVPNLPR	1333	Synpo	Isoform 3 of Synaptopodin	−2.151

IPI:IPI00404545.2	SCSVT*DTVAEQAHLPPPSV	1334	Nedd4I	Isoform 3 of E3 ubiquitin-protein	−2.113
	AYVHTTPGLPSGWEER			ligase NEDD4-like

IPI:IPI00416203.1	AVPFVPM#SYQLSQSY*Y	1335	Gtf3c1	Isoform	2 of General transcription	−2.105
	S*LGK			factor 3C polypeptide 1

IPI:IPI00355042.3	IPMPPSS*PQPR	1336	Ppp1r13l	RelA-associated inhibitor	−2.104

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAKDK	1337	Eef1b2	Elongation factor 1-beta	−2.096
	DDDIDLFGS*DDEEESEEAKK

IPI:IPI00623382.2	LMKPPSPEHQSPDTQQE	1338	Morc2a	MORC family CW-type zinc finger	−2.069
	GGEEEEAMVAR			protein 2A

IPI:IPI00125960.1	TASGSSVTSLEGTR	1339	Ndrg1	Protein NDRG1	−2.055

IPI:IPI00125960.1	TASGSSVTSLEGTR	1340	Ndrg1	Protein NDRG1	−2.055

IPI:IPI00125960.1	TASGSSVTSLEGTR{circumflex over ( )}	1341	Ndrg1	Protein NDRG1	−2.055

IPI:IPI00330121.4	GLPYADHNYGAPPPPT*PP	1342	Setd5	Isoform	2 of SET domain-containing	−2.045
	AS*PPVQTIIPR{circumflex over ( )}			protein 5

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	1343	Eif4ebp1	Eukaryotic translation initiation	−2.035
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00226701.3	ILGDHLLLDPAHELPPY*TPP	1344	Trerf1	Isoform 4 of Transcriptional-regulating	−2.03
	PMLS*PVR			factor	1

IPI:IPI00348488.5	SCS*M#ELHGEGNQEPGSP	1345	Dennd4c	DENN/MADD domain containing 4C	−2.012
	AVFAHPLER{circumflex over ( )}

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	1346	Eif4ebp1	Eukaryotic translation initiation	−2.01
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00317794.5	K@EDSDEDEDEEDEDDS	1347	Ncl	Nucleolin	−1.999
	DEDEDDEEEDEFEPPIVK@

IPI:IPI00124717.1	K@PEFS*PTLK@	1348	Orc6l	Origin recognition complex subunit 6	−1.969

IPI:IPI00380722.1	SET*PPVPPPPPYLANYPGF	1349	Bat2	Large proline-rich protein BAT2	−1.961
	PENGTPGPPISR

IPI:IPI00553798.2	SNSFS*DEREFSAPSTPTGT	1350	Ahnak	AHNAK nucleoprotein isoform 1	−1.95
	LEFAGGDAK

IPI:IPI00108389.5	LVS*PETEASEESLQFSLEK	1351	Trp53bp1	Transformation related protein 53	−1.94
	@PTTAER{circumflex over ( )}			binding protein 1

IPI:IPI00124272.1	LTTTGQVTS*PVK@	1352	Lig3	Isoform Alpha of DNA ligase 3	−1.932

IPI:IPI00406741.5	AAVGVTGNDITTPPNKEPPP	1353	Mtap4	Isoform	4 of Microtubule-associated	−1.932
	S*PEKK			protein	4

IPI:IPI00321774.2	HLLTDLPLPPELPGGDPSPP	1354	Crkrs	Isoform	2 of Cell division cycle 2-	−1.921
	DS*PEPK@			related protein kinase 7

IPI:IPI00321774.2	HLLTDLPLPPELPGGDPSPP	1355	Crkrs	Isoform	2 of Cell division cycle 2-	−1.921
	DS*PEPK@			related protein kinase 7

IPI:IPI00124914.1	SLHKPMYFFLINL	1356	Olfr521	Olfactory receptor Olfr521	−1.913
	SALDILFTTTTVPK

IPI:IPI00123881.1	VETPDVNLDQEEEIQM#ET	1357	Mcm6	DNA replication licensing factor MCM6	−1.909
	DEGQGGVNGHADS*PAPV
	NR{circumflex over ( )}

IPI:IPI00123881.1	VETPDVNLDQEEEIQM#ET	1358	Mcm6	DNA replication licensing factor MCM6	−1.909
	DEGQGGVNGHADS*PAPV
	NR

IPI:IPI00117932.2	S*PPVQPHTPVTISLGTAPS	1359	Sin3a	Isoform	1 of Paired amphipathic helix	−1.896
	LQNNQPVEFNHAINYVNK			protein Sin3a

IPI:IPI00229571.1	ISSK@SPGHM#VILNQTK@	1360	Sltm	Isoform	1 of SAFB-like transcription	−1.88
				modulator

IPI:IPI00124753.3	SLLLADSNGYTNLPDVVQP	1361	Mink1	misshapen-like kinase 1 isoform 2	−1.868
	SHSPTENS*K

IPI:IPI00117088.2	LS*VVGPPNR{circumflex over ( )}	1362	Map3k2	Mitogen-activated protein kinase kinase	−1.86
				kinase 2

IPI:IPI00127602.1	VVVADDDSEAPER{circumflex over ( )}PVNGA	1363	Slc30a4	Zinc transporter 4	−1.822
	HPALQADDDS*LLDQDLPLT
	NSQLSLK@

IPI:IPI00127602.1	VVVADDDSEAPER{circumflex over ( )}PVNGA	1364	Slc30a4	Zinc transporter	4	−1.822
	HPALQADDDS*LLDQDLPLT
	NSQLSLK@

IPI:IPI00228942.4	DLS*PGAPPAVAAAAPEER{circumflex over ( )}	1365	Hectd2	Hectd2 protein	−1.792

IPI:IPI00225062.2	EMPGSNIESSPEVEERPA	1366	Srrm2	Isoform	3 of Serine/arginine repetitive	−1.785
	VLSALDQSQSQPSK			matrix protein 2

IPI:IPI00127602.1	VVVADDDSEAPERPVNGAH	1367	Slc30a4	Zinc transporter	4	−1.782
	PALQADDDS*LLDQDLPLTN
	SQLSLK

IPI:IPI00127602.1	VVVADDDSEAPERPVNGAH	1368	Slc30a4	Zinc transporter	4	−1.782
	PALQADDDS*LLDQDLPLTN
	SQLSLK

IPI:IPI00551082.3	EK@PPFLPEEPSSSSEED	1369	Rbbp6	Isoform	2 of Retinoblastoma-binding	−1.768
	DPIPDELLCLICK@			protein 6

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	1370	Eif4ebp1	Eukaryotic translation initiation	−1.759
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00318938.6	RVALGDGVQLPPGDYSTT*	1371	Eif4ebp1	Eukaryotic translation initiation	−1.759
	PGGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00135660.5	FQHPNTDMLQEK@PSSPS	1372	Sdpr	Serum deprivation-response protein	−1.749
	PMPSST*PSPSLNLGSTEEA
	IR{circumflex over ( )}

IPI:IPI00131725.3	HQIVCINNDLSEEESDDES	1373	Npm3	Nucleoplasmin-3	−1.749
	*EEDEIKLCGILPAK

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAKD	1374	Eef1b2	Elongation factor 1-beta	−1.741
	DDDIDLFGS*DDEEESEEAKK

IPI:IPI00321774.2	HLLTDLPLPPELPGGDPS*P	1375	Crkrs	Isoform	2 of Cell division cycle 2-	−1.734
	PDSPEPK@			related protein kinase 7

IPI:IPI00420601.5	T*CDSPQNPVDFISGPVPDS	1376	Usp10	Ubiquitin carboxyl-terminal hydrolase	−1.734
	PFPR{circumflex over ( )}			10

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	1377	Npm1	Nucleophosmin	−1.718
	AES*EDEDEEDVK@

IPI:IPI00464324.2	AS*CEENLPVFIASELAK@	1378	D830031N03Rik	similar to mKIAA0754 protein	−1.718

IPI:IPI00318048.5	SSPKEEVASEPEEAASPT	1379	Nop56	Nucleolar protein 56	−1.708
	T*PK

IPI:IPI00115257.1	QVDTEEAGMVTAAT*ASNV	1380	Psip1	Isoform	1 of PC4 and SFRS1-	−1.694
	KASPK			interacting protein

IPI:IPI00317794.5	KEDSDEDEDEEDEDDSD	1381	Ncl	Nucleolin	−1.692
	EDEDDEEEDEFEPPIVK

IPI:IPI00125745.1	HAELSGS*PLK@	1382	Mta3	Isoform	1 of Metastasis associated	−1.69
				protein MTA3

IPI:IPI00461272.6	RGS*PGGVEMNVELPQQE	1383	2310047M10Rik	Putative uncharacterized protein	−1.686
	GDDDDDEDEEAAAGR

IPI:IPI00453999.2	K@PS*VPDTASPADDSFVD	1384	Nck1	non-catalytic region of tyrosine kinase	−1.682
	PGER{circumflex over ( )}			adaptor protein 1

IPI:IPI00119442.1	M#DTGEVSDIGSQGAPIVLS	1385	Exosc9	Exosome complex exonuclease RRP45	−1.674
	DSEEEEMIILEPEK@NPK@

IPI:IPI00654388.2	AENQRPAEDSALS*PGPLA	1386	Lrrfip1	Isoform	1 of Leucine-rich repeat	−1.673
	GAK			flightless-interacting protein 1

IPI:IPI00128975.3	DK@DDQEWEST*SPPK@P	1387	Dap	Death-associated protein 1	−1.669
	TVFISGVIAR{circumflex over ( )}

IPI:IPI00153375.1	SACFSPVSLS*PR{circumflex over ( )}PCSPFS	1388	Pdlim2	PDZ and LIM domain protein 2	−1.659
	TPPPTSPVALSK@

IPI:IPI00515576.5	LDGELAITKPNVSS*PSK	1389	Ehbp1	Isoform	2 of EH domain-binding	−1.659
				protein 1

IPI:IPI00169521.4	ALAEEDSEDELPSDVDFN	1390	Esf1	ESF1 homolog	−1.656
	DPYFAEEVKK

IPI:IPI00405756.8	SISGTS*TSEK@PNSMDTA	1391	Rasal2	RAS protein activator like 2	−1.654
	NTSPFK@

IPI:IPI00108508.5	ETPPTPTS*SPAEFSPK	1392	4921517D21Rik	RIKEN cDNA 4921517D21 gene	−1.652

IPI:IPI00652649.1	IRPPPELETSLTERPSSPS	1393	Als2cr4	Amyotrophic lateral sclerosis 2	−1.649
	LLR			chromosomal region candidate gene 4
				protein homolog

IPI:IPI00127415.1	LKCGSGPVHISGQHLVAVE	1394	Npm1	Nucleophosmin	−1.638
	EDAES*EDEDEEDVK

IPI:IPI00453603.1	TPVSPVKFSPGDFWGR	1395	Rps6kb1	Isoform Alpha I of Ribosomal protein	−1.635
				S6 kinase beta-1

IPI:IPI00313307.3	LSSS*DSIGPDVTDILSDIAE	1396	Med1	Isoform 4 of Mediator of RNA	−1.624
	EASK@LPSTSDDCPPIGT*P			polymerase II transcription subunit 1
	VR{circumflex over ( )}

IPI:IPI00379682.1	ANTLSHFPVECPAPPEPAQ	1397	Tbc1d1	Isoform	1 of TBC1 domain family	−1.623
	SS*PGVSQR{circumflex over ( )}			member 1

IPI:IPI00379682.1	ANTLSHFPVECPAPPEPAQ	1398	Tbc1d1	Isoform	1 of TBC1 domain family	−1.623
	S*SPGVSQR{circumflex over ( )}			member 1

IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1399	Eif4ebp1	Eukaryotic translation initiation	−1.621
	GGTLFSTT*PGGTR			factor 4E-binding protein 1

IPI:IPI00351206.5	LLK@PGEEPSEYTDEEDT	1400	Pgrmc2	Membrane-associated progesterone	−1.62
	K@			receptor component	2

IPI:IPI00225062.2	NSGPVSEVNTGFS*PEVK@	1401	Srrm2	Isoform	3 of Serine/arginine repetitive	−1.616
				matrix protein 2

IPI:IPI00112339.9	R{circumflex over ( )}LS*ENNCSLDDWEIGAGH	1402	Lima1	LIM domain and actin binding 1	−1.615
	LSSSAFNSEK@			isoform a

IPI:IPI00112339.9	R{circumflex over ( )}LS*ENNCSLDDWEIGAGH	1403	Lima1	LIM domain and actin binding 1	−1.615
	LSSSAFNSEK@			isoform a

IPI:IPI00515576.5	LDGELAITKPNVSS*PSK	1404	Ehbp1	Isoform	2 of EH domain-binding	−1.612
				protein 1

IPI:IPI00229472.5	VEEQVEDSDDEEDDDSH	1405	Rbm28	RNA-binding protein 28	−1.609
	DDEEER

IPI:IPI00229472.5	VEEQVEDSDDEEDDDSH	1406	Rbm28	RNA-binding protein 28	−1.609
	DDEEER{circumflex over ( )}

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	1407	Eif4ebp1	Eukaryotic translation initiation	−1.6
	SPTSDEPPM#QASQSQLPS			factor 4E-binding protein 1
	SPEDK

IPI:IPI00127415.1	LKCGSGPVHISGQHLVAVE	1408	Npm1	Nucleophosmin	−1.599
	EDAES*EDEDEEDVK

IPI:IPI00453837.2	KLPPPPPQAPPEEENES*EP	1409	Kdm1	Lysine-specific histone demethylase 1	−1.587
	EEPSGVEGAAFQSR

IPI:IPI00319933.4	DGTAPPPQSPSSPGSGQD	1410	Pacsin3	Protein kinase C and casein kinase II	−1.565
	EDWS*DEESPR			substrate protein 3

IPI:IPI00125662.2	S*PQVPAAQQMLNFPEK	1411	Smarcc1	Isoform 1 of SWI/SNF complex subunit	−1.554
				SMARCC1

IPI:IPI00230435.1	SLTM#VEDNEDDDEDGEEL	1412	Lmna	Isoform C2 of Lamin-A/C	−1.546
	LHHHR{circumflex over ( )}VS*GSR{circumflex over ( )}

IPI:IPI00228748.1	LDR{circumflex over ( )}AS*PDLWPEQLPGVAE	1413	Lin52;	Protein lin-52 homolog	−1.545
	FAASFK@		Gm7020

IPI:IPI00622364.4	LPPPFPGMDPESEGAAGAS	1414	Snx1	Sorting nexin-1	−1.535
	EPEAGDSDTEGEDIFTGAA
	AATK@PQSPK@

IPI:IPI00117689.1	GEATDLLRGSSPDVHTLL	1415	Ptrf	Polymerase I and transcript release	−1.532
	EITEESDAVLVDK			factor

IPI:IPI00117689.1	GEATDLLR{circumflex over ( )}GSSPDVHTLL	1416	Ptrf	Polymerase I and transcript release	−1.532
	EITEESDAVLVDK@			factor

IPI:IPI00606555.2	KPEPEAGQAEPLS*PRPPP	1417	Rreb1	Isoform	3 of Ras-responsive element-	−1.526
	CPTLSVTVEPK			binding protein 1

IPI:IPI00606555.2	KPEPEAGQAEPLS*PRPPP	1418	Rreb1	Isoform 3 of Ras-responsive element-	−1.526
	CPTLSVTVEPK			binding protein 1

IPI:IPI00380394.5	IEEPVSM#EMDNHLSDKDE	1419	Ints3	Integrator complex subunit 3	−1.525
	SCYDNAEAAFS*DDEEDLN
	SK

IPI:IPI00229571.1	ISSK@SPGHM#VILNQTK@	1420	Sltm	Isoform	1 of SAFB-like transcription	−1.524
				modulator

IPI:IPI00551082.3	EKPPFLPEEPSSSSEEDD	1421	Rbbp6	Isoform	2 of Retinoblastoma-binding	−1.522
	PIPDELLCLICK			protein	6

IPI:IPI00468896.7	KGELMENDQDAMEYSSE	1422	Ddx46	Isoform	1 of Probable ATP-dependent	−1.517
	EEEVDLQTALTGYQTK			RNA helicase DDX46

IPI:IPI00280250.5	VGES*SEDVALEEETIYENE	1423	Sh3pxd2a	Isoform	1 of SH3 and PX domain-	−1.513
	GFR{circumflex over ( )}PYTEDTLSAR{circumflex over ( )}			containing protein 2A

IPI:IPI00280250.5	VGES*SEDVALEEETIYENE	1424	Sh3pxd2a	Isoform	1 of SH3 and PX domain-	−1.513
	GFRPYTEDTLSAR			containing protein 2A

IPI:IPI00664670.4	LGS*FGSITR	1425	Flnc	Isoform	1 of Filamin-C	−1.512

IPI:IPI00117929.1	STEESLSEDAFTESELS*PIR	1426	Oxr1	Isoform	2 of Oxidation resistance	−1.51
	{circumflex over ( )}EELLSSEPR{circumflex over ( )}			protein 1

IPI:IPI00117929.1	STEESLSEDAFTESELS*PIR	1427	Oxr1	Isoform	2 of Oxidation resistance	−1.51
	{circumflex over ( )}EELLSSEPR{circumflex over ( )}			protein 1

IPI:IPI00664670.4	LGS*FGSITR{circumflex over ( )}	1428	Flnc	Isoform	1 of Filamin-C	−1.509

IPI:IPI00130920.1	ASLSPMDEPVPDSESPVE	1429	Mtap1b	Microtubule-associated protein 1B	−1.509
	K@

IPI:IPI00113362.2	NSNSYGIPEPAHAY*AQPQT	1430	Crkl	Crk-like protein	−1.508
	TTPLPTVASTPGAAINPLPS
	TQNGPVFAK

IPI:IPI00331334.3	VSSAPIPCPS*PSPAPSAVP	1431	Bag3	BAG family molecular chaperone	−1.508
	S*PPK			regulator	3

IPI:IPI00331209.1	NSS*FVLPK	1432	Rin2	Isoform	1 of Ras and Rab interactor 2	−1.504

IPI:IPI00225062.2	GHTQTWPDTSSPEVMQT*Q	1433	Srrm2	Isoform	3 of Serine/arginine repetitive	−1.504
	VES*PLLQSK			matrix protein	2

IPI:IPI00664670.4	LGS*FGSITR{circumflex over ( )}	1434	Flnc	Isoform	1 of Filamin-C	−1.502

IPI:IPI00664670.4	LGS*FGSITR	1435	Flnc	Isoform	1 of Filamin-C	−1.502

IPI:IPI00664670.4	LGS*FGSITR	1436	Flnc	Isoform	1 of Filamin-C	−1.502

IPI:IPI00664670.4	LGS*FGSITR	1437	Flnc	lsoform	1 of Filamin-C	−1.497

IPI:IPI00126006.6	TQT*PPLGQTPQLGLK@	1438	Eif4g2	Isoform 1 of Eukaryotic translation	−1.492
				initiation factor 4 gamma 2

IPI:IPI00113798.2	ATWGDGGDNS*PSNVVSK	1439	Snap23	Synaptosomal-associated protein	−1.49

IPI:IPI00664670.4	LGS*FGSITR{circumflex over ( )}	1440	Flnc	Isoform	1 of Filamin-C	−1.483

IPI:IPI00169771.1	EDGIDAVEVAADR{circumflex over ( )}PGS*PR{circumflex over ( )}	1441	BC031781	UPF0667 protein C1orf55 homolog	−1.482

IPI:IPI00169806.3	GMEGLIS*PTEAVGNSCGA	1442	Nek9	Serine/threonine-protein kinase Nek9	−1.478
	SSSCPGWLR{circumflex over ( )}

IPI:IPI00117932.2	S*PPVQPHTPVTISLGTAPS	1443	Sin3a	Isoform 1 of Paired amphipathic helix	−1.477
	LQNNQPVEFNHAINYVNK@			protein Sin3a

IPI:IPI00225062.2	NSGPVSEVNTGFS*PEVK@	1444	Srrm2	Isoform	3 of Serine/arginine repetitive	−1.475
				matrix protein 2

IPI:IPI00128062.1	RGQEELEVPVDPLPS*SPA	1445	Snx15	Sorting nexin-15	−1.472
	QEALDLLFSCDSTEEASSSL
	AR

IPI:IPI00224570.3	GVNFAEEPMRSDSENGE	1446	Prkar2b	cAMP-dependent protein kinase type II-	−1.468
	EEEAAEAGAFNAPVINR			beta regulatory subunit

IPI:IPI00380722.1	SET*PPVPPPPPYLANYPGF	1447	Bat2	Large proline-rich protein BAT2	−1.463
	PENGTPGPPISR{circumflex over ( )}

IPI:IPI00330695.5	GLLAQDLQAESSPPAS*PLL	1448	Fam129b	Niban-like protein 1	−1.46
	NGAPVQESSQPVAVPEAS*
	PPASPLR{circumflex over ( )}

IPI:IPI00750546.1	GGGQSS*PQEEPTWK@	1449	Cwc22	Novel protein	−1.452

IPI:IPI00129356.1	SGS*GMSVISSSSVDQR	1450	Itsn1	Isoform 1 of Intersectin-1	−1.451

IPI:IPI00753701.3	SPTLGEDSDTEVDEDHKP	1451	Mdc1	mediator of DNA damage checkpoint 1	−1.447
	GFADS*ETDVEEER

IPI:IPI00137730.7	VLTPTQVMNR{circumflex over ( )}PSS*ISWDG	1452	Pebp1	Phosphatidylethanolamine-binding	−1.431
	LDPGK@			protein 1

IPI:IPI00123802.5	NIQQDNSEAGTQPQVQTDG	1453	Hsph1	Isoform HSP105-alpha of Heat shock	−1.428
	QQTSQSPPS*PELTSEESK			protein 105 kDa
	@TPDADK@

IPI:IPI00421179.1	EATLPPVS*PPK@	1454	Eif4g1	Isoform 1 of Eukaryotic translation	−1.421
				initiation factor 4 gamma 1

IPI:IPI00421179.1	EATLPPVS*PPK	1455	Eif4g1	Isoform 1 of Eukaryotic translation	−1.421
				initiation factor 4 gamma 1

IPI:IPI00318006.5	R{circumflex over ( )}IS*WPENSFDFVSK@	1456	Serhl	Serine hydrolase-like protein	−1.411

IPI:IPI00318006.5	R{circumflex over ( )}IS*WPENSFDFVSK@	1457	Serhl	Serine hydrolase-like protein	−1.411

IPI:IPI00125501.1	KAEEAT*PVTALR	1458	Epb4.1l3	Isoform	1 of Band 4.1-like protein 3	−1.408

IPI:IPI00405665.6	K@IIETM#SS*PK@	1459	Kif20b	Isoform 1 of M-phase phosphoprotein 1	−1.399

IPI:IPI00405665.6	KIIETM#SS*PK	1460	Kif20b	Isoform	1 of M-phase phosphoprotein 1	−1.399

IPI:IPI00119442.1	M#DTGEVSDIGSQGAPIVLS	1461	Exosc9	Exosome complex exonuclease RRP45	−1.396
	DSEEEEMIILEPEK@NPK@

IPI:IPI00121760.5	LKTEEGEIVYS*AEESENR	1462	Hnrpll	Isoform	1 of Heterogeneous nuclear	−1.387
				ribonucleoprotein L-like

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAKD	1463	Eef1b2	Elongation factor 1-beta	−1.381
	DDDIDLFGS*DDEEESEEAK

IPI:IPI00664670.4	LGS*FGSITR{circumflex over ( )}	1464	Flnc	Isoform	1 of Filamin-C	−1.373

IPI:IPI00664670.4	LGS*FGSITR	1465	Flnc	Isoform	1 of Filamin-C	−1.373

IPI:IPI00125348.2	AEEQLPPLLSPPS*PSTPHSR	1466	Mprip	Isoform	1 of Myosin phosphatase Rho-	−1.371
				interacting protein

IPI:IPI00120095.2	DFTKPQDGDIIAPLIT*PLK	1467	Samhd1	SAM domain and HD domain-	−1.371
				containing protein 1

IPI:IPI00756765.3	ALVIQESESPPSPPPS	1468	Ehmt2	Euchromatic histone lysine N-	−1.368
	PDRR			methyltransferase	2

IPI:IPI00135708.1	RKPVLPALTINPTIAEGPS*P	1469	Map2k2	Dual specificity mitogen-activated	−1.365
	TSEGASEANLVDLQK			protein kinase kinase 2

IPI:IPI00317794.5	KEDSDEDEDEEDEDDSD	1470	Ncl	Nucleolin	−1.364
	EDEDDEEEDEFEPPIVK

IPI:IPI00330695.5	GLLAQDLQAESSPPASPLL	1471	Fam129b	Niban-like protein 1	−1.36
	NGAPVQESSQPVAVPEAS
	*PPASPLR{circumflex over ( )}

IPI:IPI00330695.5	GLLAQDLQAESSPPASPL	1472	Fam129b	Niban-like protein 1	−1.36
	LNGAPVQESSQPVAVPEAS
	*PPASPLR

IPI:IPI00453800.4	K@PSSSPDLWK@VSPDQ	1473	Zfp828	Zinc finger protein 828	−1.358
	R{circumflex over ( )}

IPI:IPI00135443.2	AS*PITNDGEDEFVPSDGLD	1474	Top2b	DNA topoisomerase 2-beta	−1.357
	KDEYAFSSGK

IPI:IPI00227267.2	EVENEQTPVS*EPEEEKGS	1475	Epb4.1l2	Putative uncharacterized protein	−1.357
	QPGPPVER

IPI:IPI00649691.3	LASEDAALVDDDEES*DTPA	1476	Cc2d1b	Coiled-coil and C2 domain-containing	−1.353
	QAPLAK@			protein 1B

IPI:IPI00340860.5	TAS*EGSEAETPEAPK@	1477	Larp7	Isoform	1 of La-related protein 7	−1.347

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAKD	1478	Eef1b2	Elongation factor 1-beta	−1.336
	DDDIDLFGS*DDEEESEEAK

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAKD	1479	Eef1b2	Elongation factor 1-beta	−1.336
	DDDIDLFGS*DDEEESEEAK

IPI:IPI00750546.1	GGGQSS*PQEEPTWK	1480	Cwc22	Novel protein	−1.327

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	1481	Npm1	Nucleophosmin	−1.324
	AES*EDEDEEDVK

IPI:IPI00123871.2	VHEPPR{circumflex over ( )}EDTVPPK@PVPP	1482	Ncor2	nuclear receptor co-repressor 2	−1.31
	VPPPTQHLQPEGDVSQQS
	GGS*PR{circumflex over ( )}

IPI:IPI00458583.3	AKS*PQPPVEEEDEHFDDT	1483	Hnrnpu	Putative uncharacterized protein	−1.308
	VVCLDTYNCDLHFK

IPI:IPI00124959.1	GPIQTLGHTDESASDKGPT	1484	Mki67	Ki-67 protein	−1.306
	QMPCNSLQPEQVDSFQSS*
	PR

IPI:IPI00664670.4	LGS*FGSITR	1485	Flnc	Isoform	1 of Filamin-C	−1.29

IPI:IPI00664670.4	LGS*FGSITR{circumflex over ( )}	1486	Flnc	Isoform	1 of Filamin-C	−1.289

IPI:IPI00664670.4	LGS*FGSITR{circumflex over ( )}	1487	Flnc	Isoform	1 of Filamin-C	−1.289

IPI:IPI00664670.4	LGS*FGSITR	1488	Flnc	Isoform	1 of Filamin-C	−1.289

IPI:IPI00664670.4	LGS*FGSITR	1489	Flnc	Isoform	1 of Filamin-C	−1.289

IPI:IPI00270767.3	M#K@ESETFSDSS*PIEIIDE	1490	Rtn4	Isoform	2 of Reticulon-4	−1.286
	FPTFVSAK@

IPI:IPI00318048.5	SSPKEEVASEPEEAASPT	1491	Nop56	Nucleolar protein 56	−1.284
	TPK

IPI:IPI00276222.1	LSIMTSENHLNNSDKEVDEV	1492	Fermt2	Fermitin family homolog 2	−1.279
	DAALS*DLEITLEGGK

IPI:IPI00475055.3	RCPSTDPEEAVEDAEGPS	1493	Zfhx3	AT motif binding factor 1	−1.274
	EASADPEELAK

IPI:IPI00664670.4	LGS*FGSITR{circumflex over ( )}	1494	Flnc	Isoform	1 of Filamin-C	−1.273

IPI:IPI00664670.4	LGS*FGSITR	1495	Flnc	Isoform	1 of Filamin-C	−1.273

IPI:IPI00664670.4	LGS*FGSITR{circumflex over ( )}	1496	Flnc	Isoform	1 of Filamin-C	−1.273

IPI:IPI00664670.4	LGS*FGSITR	1497	Flnc	Isoform	1 of Filamin-C	−1.273

IPI:IPI00664670.4	LGS*FGSITR{circumflex over ( )}	1498	Flnc	Isoform	1 of Filamin-C	−1.273

IPI:IPI00664670.4	LGS*FGSITR	1499	Flnc	Isoform	1 of Filamin-C	−1.273

IPI:IPI00664670.4	LGS*FGSITR{circumflex over ( )}	1500	Flnc	Isoform	1 of Filamin-C	−1.272

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	1501	Npm1	Nucleophosmin	−1.271
	AES*EDEDEEDVK@

IPI:IPI00121277.1	VAAAAGSGPSPPCSPGH	1502	Pi4k2a	Phosphatidylinositol 4-kinase type 2-	−1.266
	DR			alpha

IPI:IPI00121277.1	VAAAAGSGPSPPCSPGH	1503	Pi4k2a	Phosphatidylinositol 4-kinase type 2-	−1.266
	DR{circumflex over ( )}			alpha

IPI:IPI00317794.5	K@EDSDEDEDEEDEDDS	1504	Ncl	Nucleolin	−1.255
	DEDEDDEEEDEFEPPIVK@

IPI:IPI00270767.3	VVKEDGVM#S*PEK	1505	Rtn4	Isoform	2 of Reticulon-4	−1.252

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	1506	Eif4ebp1	Eukaryotic translation initiation	−1.238
	SPTSDEPPM#QASQSQLPS			factor 4E-binding protein 1
	SPEDK

IPI:IPI00114352.1	VNHEPEPASGAS*PGATIPK	1507	Bin1	Isoform	1 of Myc box-dependent-	−1.237
	@S*PSQLR{circumflex over ( )}			interacting protein 1

IPI:IPI00114352.1	VNHEPEPASGAS*PGATIPK	1508	Bin1	Isoform 1 of Myc box-dependent-	−1.237
	S*PSQLR			interacting protein	1

IPI:IPI00110435.2	TPS*PEPVDK@DFYSEFGD	1509	Nisch	Isoform 1 of Nischarin	−1.229
	K@

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	1510	Npm1	Nucleophosmin	−1.226
	AES*EDEDEEDVK@

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	1511	Npm1	Nucleophosmin	−1.226
	AES*EDEDEEDVK@

IPI:IPI00132386.1	ANS*TSDSMFTETPSPVLK@	1512	Cdc42ep3	Cdc42 effector protein 3	−1.225

IPI:IPI00131999.2	FELLPTPPLSPS*RR	1513	Myc	myc proto-oncogene protein	−1.219

IPI:IPI00282957.4	EPAAPASPAPSPVPSPT*P	1514	Mtap7d1	Isoform 1 of MAP7 domain-containing	−1.217
	AQPQK			protein	1

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	1515	Eif4ebp1	Eukaryotic translation initiation	−1.212
	SPTSDEPPMQASQSQLPSS			factor 4E-binding protein 1
	PEDK

IPI:IPI00226220.2	TFS*LDAAPADHSLGPSR{circumflex over ( )}	1516	Rtkn	Isoform 1 of Rhotekin	−1.207

IPI:IPI00318938.6	NSPVAKTPPKDLPAIPGVT	1517	Eif4ebp1	Eukaryotic translation initiation	−1.203
	SPTSDEPPM#QASQSQLPS			factor 4E-binding protein 1
	SPEDK

IPI:IPI00664670.4	LGSFGSITR{circumflex over ( )}	1518	Flnc	Isoform	1 of Filamin-C	−1.194

IPI:IPI00473912.4	NSTHSNLHTSLGNSVWG	1519	Gigyf2	Isoform 1 of PERQ amino acid-rich with	−1.193
	SINTGPSNQWASELVSSI			GYF domain-containing protein 2
	WSNADTK

IPI:IPI00227380.1	TRS*EPLPPSATASPLLAPL	1520	Hdac7	Isoform	4 of Histone deacetylase 7	−1.178
	QPR

IPI:IPI00119442.1	M#DTGEVSDIGSQGAPIVLS	1521	Exosc9	Exosome complex exonuclease RRP45	−1.177
	DSEEEEMIILEPEK@NPK@

IPI:IPI00119442.1	M#DTGEVSDIGSQGAPIVLS	1522	Exosc9	Exosome complex exonuclease RRP45	−1.177
	DSEEEEMIILEPEKNPK

IPI:IPI00119442.1	MDTGEVSDIGSQGAPIVLS*	1523	Exosc9	Exosome complex exonuclease RRP45	−1.177
	DS*EEEEM#IILEPEKNPK

IPI:IPI00221882.1	DHNS*EDEDEDKYADDIDM	1524	Slu7	Pre-mRNA-splicing factor SLU7	−1.167
	#PGQNFDSK

IPI:IPI00675346.2	R{circumflex over ( )}QSGT*DLQEDVIVR{circumflex over ( )}	1525	Myo9a	Isoform 2 of Myosin-IXa	−1.166

IPI:IPI00125319.1	TTS*FAESCKPVQQPSAFG	1526	Gsk3b	Glycogen synthase kinase-3 beta	−1.165
	SMK

IPI:IPI00654192.2	SLS*FSEPQQPPPTVK	1527	Zfp395	zinc finger protein 395	−1.165

IPI:IPI00128975.3	DKDDQEWEST*SPPKPTVFI	1528	Dap	Death-associated protein 1	−1.165
	SGVIAR

IPI:IPI00330897.1	SPVVITIDSDS*	1529	Topors	E3 ubiquitin-protein ligase Topors	−1.161
	DGESEVK@

IPI:IPI00330133.2	INDELEGLGLEGGS*EGEAPR	1530	Bend3	BEN domain-containing protein 3	−1.159

IPI:IPI00330133.2	INDELEGLGLEGGS*EGEAPR	1531	Bend3	BEN domain-containing protein 3	−1.159

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	1532	Npm1	Nucleophosmin	−1.157
	AES*EDEDEEDVK@

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	1533	Npm1	Nucleophosmin	−1.157
	AES*EDEDEEDVK

IPI:IPI00314749.2	ISENYS*DKSDVENADESSS	1534	Slc4a4	Isoform	1 of Electrogenic sodium	−1.156
	SILKPLISPAAER			bicarbonate cotransporter 1

IPI:IPI00123709.1	GPSEAPQEAEAEEGATS*D	1535	Akap12	Isoform 1 of A-kinase anchor protein 12	−1.154
	GEK

IPI:IPI00321774.2	HLLTDLPLPPELPGGDPS*P	1536	Crkrs	Isoform	2 of Cell division cycle 2-	−1.15
	PDS*PEPK			related protein kinase 7

IPI:IPI00420601.5	SDLIEDEELEDTGKGS*EDE	1537	Usp10	Ubiquitin carboxyl-terminal hydrolase	−1.149
	WEQVGPK			10

IPI:IPI00421179.1	EATLPPVS*PPK@	1538	Eif4g1	Isoform 1 of Eukaryotic translation	−1.146
				initiation factor 4 gamma 1

IPI:IPI00269136.2	KGLVAASGSDSEDEDSM	1539	Tgif1	Isoform 1 of Homeobox protein TGIF1	−1.143
	DSPLDLSSSAASGK

IPI:IPI00221581.1	TGS*ESSQTGASATSGR	1540	Eif4b	Eukaryotic translation initiation	−1.142
				factor 4B

IPI:IPI00756765.3	ALVIQESESPPSPPPSPDR	1541	Ehmt2	Euchromatic histone lysine N-	−1.141
	{circumflex over ( )}R{circumflex over ( )}			methyltransferase 2

IPI:IPI00137668.1	M#LPHAPGVQM#QAIPEDA	1542	Hdac2	Histone deacetylase	2	−1.123
	VHEDS*GDEDGEDPDKR

IPI:IPI00225062.2	IHTTALTGQS*PPLASGHQG	1543	Srrm2	Isoform	3 of Serine/arginine repetitive	−1.123
	EGDAPSVEPGATNIQQPSS*			matrix protein 2
	PAPSTK@

IPI:IPI00225062.2	IHTTALTGQS*PPLASGHQG	1544	Srrm2	Isoform 3 of Serine/arginine repetitive	−1.123
	EGDAPSVEPGATNIQQPSS*			matrix protein 2
	PAPSTK

IPI:IPI00225062.2	IHTTALTGQSPPLASGHQG	1545	Srrm2	Isoform	3 of Serine/arginine repetitive	−1.123
	EGDAPS*VEPGATNIQQPSS			matrix protein 2
	*PAPSTK

IPI:IPI00339428.9	MSSHT*ETSSFLQTLTGR{circumflex over ( )}	1546	Dock7	Isoform 2 of Dedicator of cytokinesis	−1.12
				protein 7

IPI:IPI00339428.9	MS*SHTETSSFLQTLTGR{circumflex over ( )}	1547	Dock7	Isoform 2 of Dedicator of cytokinesis	−1.12
				protein 7

IPI:IPI00339428.9	MS*SHTETSSFLQTLTGR	1548	Dock7	Isoform 2 of Dedicator of cytokinesis	−1.12
				protein 7

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	1549	Npm1	Nucleophosmin	−1.119
	AES*EDEDEEDVK@

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	1550	Npm1	Nucleophosmin	−1.119
	AES*EDEDEEDVK@

IPI:IPI00331361.2	S*PSLLQSGVK	1551	Mybbp1a	Myb-binding protein 1A	−1.116

IPI:IPI00309059.7	STSPIIGSPPVR{circumflex over ( )}	1552	Patl1	Protein PAT1 homolog 1	−1.115

IPI:IPI00339428.9	MSSHTETS*SFLQTLTGR{circumflex over ( )}	1553	Dock7	Isoform 2 of Dedicator of cytokinesis	−1.114
				protein 7

IPI:IPI00123747.1	LPS*M#GDQEPPGQEK	1554	Slc7a11	Cystine/glutamate transporter	−1.114

IPI:IPI00322095.4	IDEPNTPYHNMIGDDEDAYS	1555	Ppp1r2	Putative uncharacterized protein	−1.107
	DS*EGNEVM#TPDILAK

IPI:IPI00135379.3	LSEGEPGPVAAGEQLSEHP	1556	Myo9b	Isoform 1 of Myosin-IXb	−1.106
	VEDPESLGVEAETWMNKS*
	PDGMS*PK

IPI:IPI00125319.1	TTS*FAESCKPVQQPSAFG	1557	Gsk3b	Glycogen synthase kinase-3 beta	−1.105
	SMK

IPI:IPI00125319.1	TTS*FAESCK@PVQQPSAF	1558	Gsk3b	Glycogen synthase kinase-3 beta	−1.105
	GSMK@

IPI:IPI00125319.1	TTS*FAESCKPVQQPSAFG	1559	Gsk3b	Glycogen synthase kinase-3 beta	−1.103
	SMK

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAKD	1560	Eef1b2	Elongation factor 1-beta	−1.102
	DDDIDLFGS*DDEEESEEAKK

IPI:IPI00331361.2	SPAPSNPTLSPSTPAK@T	1561	Mybbp1a	Myb-binding protein 1A	−1.101
	PK@

IPI:IPI00331361.2	SPAPSNPTLSPSTPAKTPK	1562	Mybbp1a	Myb-binding protein 1A	−1.101

IPI:IPI00668709.2	SSS*APNVHINTI	1563	Braf	B-Raf protein	−1.097
	EPVNIDDLIR

IPI:IPI00462466.6	K@LDES*PVLK@PEFIGHD	1564	Setd2	SET domain containing 2	−1.096
	GR{circumflex over ( )}

IPI:IPI00462466.6	KLDES*PVLKPEFIGHDGR	1565	Setd2	SET domain containing 2	−1.096

IPI:IPI00108389.5	LLDGPTGSSEEEEEFLEIP	1566	Trp53bp1	Transformation related protein 53	−1.094
	PFNK			binding protein	1

IPI:IPI00668709.2	SSS*APNVHINTIEP	1567	Braf	B-Raf protein	−1.088
	VNIDDLIR

IPI:IPI00225062.2	DGSGTPSRHSLSGSS*PG	1568	Srrm2	Isoform	3 of Serine/arginine repetitive	−1.081
	M#KDTPQT*PSR			matrix protein	2

IPI:IPI00135971.1	VQIPVSHPDPEPVS*DNEDD	1569	Tjp1	Tight junction protein ZO-1	−1.08
	S*YDEEVHDPR

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	1570	Npm1	Nucleophosmin	−1.077
	AES*EDEDEEDVK

IPI:IPI00111169.1	HIVSNDSSDS*DDEAQGPK@	1571	Pla2g4a	Cytosolic phospholipase A2	−1.072

IPI:IPI00225670.1	EVHDELEDLPS*PPPPLSPP	1572	Gphn	Gephyrin	−1.07
	PT*TSPHK

IPI:IPI00225670.1	EVHDELEDLPSPPPPLSP	1573	Gphn	Gephyrin	−1.07
	PPTTSPHK@

IPI:IPI00123709.1	AEDS*GAEQLASEIEPSR	1574	Akap12	Isoform 1 of A-kinase anchor protein 12	−1.067

IPI:IPI00123709.1	AEDS*GAEQLASEIEPSR	1575	Akap12	Isoform 1 of A-kinase anchor protein 12	−1.067

IPI:IPI00454109.2	S*PPACSSSSSSLFSAVVAR{circumflex over ( )}	1576	Erf	ETS domain-containing transcription	−1.066
				factor ERF

IPI:IPI00313517.1	GPPS*EDEGMDIHFEEGVLS	1577			−1.066
	PSAADMR{circumflex over ( )}PEPPNSLDLNG
	SHPR{circumflex over ( )}

IPI:IPI00313517.1	GPPS*EDEGMDIHFEEGVLS	1578			−1.066
	PSAADMRPEPPNSLDLNGS
	HPR

IPI:IPI00311375.6	S*GITSLLFGEDDLEALK	1579	Sgsm3	small G protein signaling modulator 3	−1.061

IPI:IPI00311490.4	VLSDSEEEEKDADVPGTS	1580	Phip	PH-interacting protein	−1.059
	TR

IPI:IPI00553798.2	GHYEVT*GSDDEAGKLQGS	1581	Ahnak	AHNAK nucleoprotein isoform 1	−1.057
	GVSLASK

IPI:IPI00553798.2	GHYEVTGS*DDEAGK@LQ	1582	Ahnak	AHNAK nucleoprotein isoform 1	−1.057
	GSGVSLASK@

IPI:IPI00336281.1	RDSICS*SVSMESSLAEPQD	1583	Golga3	Isoform 1 of Golgin subfamily A	−1.056
	ELLQILK			member 3

IPI:IPI00229859.1	ALENGEADEPSFSDPEDF	1584	Eif3b	Eif3b protein	−1.053
	VDDVSEEELLGDVLK

IPI:IPI00668709.2	DRSSS*APNVHINTIEPVNID	1585	Braf	B-Raf protein	−1.048
	DLIR

IPI:IPI00668709.2	DR{circumflex over ( )}SSS*APNVHINTIEPVNI	1586	Braf	B-Raf protein	−1.048
	DDLIR{circumflex over ( )}

IPI:IPI00125319.1	T*TSFAESCKPVQQPSAFG	1587	Gsk3b	Glycogen synthase kinase-3 beta	−1.047
	SMK

IPI:IPI00135660.5	RGNNSAVGSNADLTIEED	1588	Sdpr	Serum deprivation-response protein	−1.046
	EEEEPVALQQAQQVR

IPI:IPI00339428.9	MSS*HTETSSFLQTLTGR	1589	Dock7	Isoform 2 of Dedicator of cytokinesis	−1.042
				protein 7

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	1590	Npm1	Nucleophosmin	−1.039
	AES*EDEDEEDVK

IPI:IPI00115660.1	GPILAT*PGK@	1591	Tcof1	Treacle protein	−1.038

IPI:IPI00115660.1	GPILAT*PGK	1592	Tcof1	Treacle protein	−1.038

IPI:IPI00115660.1	GPILAT*PGK	1593	Tcof1	Treacle protein	−1.038

IPI:IPI00115660.1	GPILAT*PGK@	1594	Tcof1	Treacle protein	−1.038

IPI:IPI00230704.4	KES*APQVLLPEEEK	1595	Arhgef7	Isoform C of Rho guanine nucleotide	−1.036
				exchange factor 7

IPI:IPI00320422.3	LQEEGGS*EEEEAGNPSED	1596	Pwp1	Periodic tryptophan protein 1 homolog	−1.035
	GM#QSGPTQAPPR

IPI:IPI00270767.3	VVKEDGVM#S*PEK	1597	Rtn4	Isoform	2 of Reticulon-4	−1.035

IPI:IPI00331306.7	ARQPSQADTGEEDSDEDY*	1598	Sh3bp2	SH3-domain binding protein 2 isoform c	−1.034
	EKVPLPNSVFVNTTESCEV
	ER

IPI:IPI00137864.1	YDLSNQDHIMDAPPLS*PFP	1599	Rbl1	Isoform Long of Retinoblastoma-like	−1.03
	HIK@			protein 1

IPI:IPI00137864.1	YDLSNQDHIMDAPPLS*PFP	1600	Rbl1	Isoform Long of Retinoblastoma-like	−1.03
	HIK			protein	1

IPI:IPI00320208.3	YGPSSVEDTTGSGAADAK	1601	Eef1b2	Elongation factor 1-beta	−1.028
	@DDDDIDLFGS*DDEEESE
	EAK@K@

IPI:IPI00404545.2	SLSSPTVTLSAPLEGAK	1602	Nedd4l	Isoform 3 of E3 ubiquitin-protein	−1.027
				ligase NEDD4-like

IPI:IPI00127764.2	VIEDEDGAAAAATVSNSEET	1603	Pcm1	Isoform	1 of Pericentriolar material 1	−1.024
	PIIENHNS*PQPISDVSAVPC			protein
	PR{circumflex over ( )}

IPI:IPI00131884.1	TTPAPSPGS*ANESFFAPS	1604	Mid1ip1	Mid1-interacting protein 1	−1.024
	R{circumflex over ( )}

IPI:IPI00131884.1	TTPAPSPGS*ANESFFAPS	1605	Mid1ip1	Mid1-interacting protein 1	−1.024
	R{circumflex over ( )}

IPI:IPI00131884.1	TTPAPSPGS*ANESF	1606	Mid1ip1	Mid1-interacting protein 1	−1.024
	FAPSR

IPI:IPI00400381.9	LGSTGSQPNSEAEPGPEH	1607	Phactr4	Isoform	1 of Phosphatase and actin	−1.02
	APK			regulator	4

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	1608	Npm1	Nucleophosmin	−1.018
	AES*EDEDEEDVK

IPI:IPI00756765.3	ALVIQESESPPSPPPSPDR	1609	Ehmt2	Euchromatic histone lysine N-	−1.017
	{circumflex over ( )}R{circumflex over ( )}			methyltransferase 2

IPI:IPI00136917.5	TDQEGLNASQPT*PPPLPK@	1610	C230081A13Rik	Tyrosine-protein kinase-protein kinase	−1.016

IPI:IPI00127415.1	CGSGPVHISGQHLVAVEED	1611	Npm1	Nucleophosmin	−1.015
	AES*EDEDEEDVK

IPI:IPI00453673.4	RAS*DGGANIQLHAQQLLK	1612	BC033915	serine/threonine-protein kinase QSK	−1.015

IPI:IPI00453837.2	KLPPPPPQAPPEEENES*EP	1613	Kdm1	Lysine-specific histone demethylase 1	−1.014
	EEPSGVEGAAFQSR

IPI:IPI00227900.1	FKGPGDTSNFDDYEEEEIR	1614	Prkaca	Isoform 2 of cAMP-dependent protein	−1.01
	VS*INEK			kinase catalytic subunit alpha

IPI:IPI00125662.2	VDPTYGLESSCIAGTGPDE	1615	Smarcc1	Isoform 1 of SWI/SNF complex subunit	−1.007
	PEK@LEGS*EEEK@M#ETD			SMARCC1
	PDGQQPEK@

IPI:IPI00677756.1	KLS*FSMPR	1616	Ahnak2	Putative uncharacterized protein	−1.006

IPI:IPI00395213.1	LDFLPEMMVDHCS*LNSSP	1617	Cugbp1	Isoform	4 of CUG-BP- and ETR-3-like	−1.002
	VSK@K@			factor 1

Table 8. Gene names and sequences of the phosphopeptides whose intensities decrease after Ku-0063794 treatment. Note that the light cells were treated with rapamycin and serve as controls whereas the heavy cells were treated with a combination of rapamycin and Ku-0063794.


Table 9(a)

			SEQ ID	Gene
ID	Reference	Ascore Seq	NO	Name	Annotation	Class

1	IPI:IPI00121418.1	DGEGPDNLEPACPLSLPLQ	1618	Rb1	Retinoblastoma-associated	1
		GNHTAADMYLSPLRSPK			protein

2	IPI:IPI00221581.1	TGSESSQTGASATSGR	1619	Eif4b	Eukaryotic translation initiation	1
					factor 4B

3	IPI:IPI00929786.1	TASISSSPSEGTPAVGSY	1620	Larp1	Isoform	1 of La-related protein 1	1
		GCTPQSLPK@

4	IPI:IPI00929786.1	TASISSSPSEGTPAVGSY*	1621	Larp1	Isoform	1 of La-related protein 1	1
		GCT*PQSLPK@

5	IPI:IPI00929786.1	TASISSSPSEGTPAVGS*Y	1622	Larp1	Isoform	1 of La-related protein 1	1
		GCTPQSLPK@

6	IPI:IPI00317401.6	AESPETSAVESTQSTPQK@	1623	Pds5b	Isoform	1 of Sister chromatid	1
					cohesion protein PDS5 homolog B

7	IPI:IPI00107958.1	TSDIFGS*PVTATAPLAHPN	1624	Hn1l	Hematological and neurological	1
		K@PK@			expressed 1-like protein

8	IPI:IPI00108454.2	R{circumflex over ( )}LSSLR{circumflex over ( )}	1625	Rps6	29 kDa protein	1

9	IPI:IPI00225062.2	R{circumflex over ( )}SSSELSPEVVEK@	1626	Srrm2	Isoform	3 of Serine/arginine	1
					repetitive matrix protein 2

10	IPI:IPI00761759.1	SSSGS*EHSTEGSVSLGD	1627	Larp4	Putative uncharacterized protein	1
		GPLSR{circumflex over ( )}

11	IPI:IPI00761759.1	SSSGSEHST*EGSVSLGD	1628	Larp4	Putative uncharacterized protein	1
		GPLSR{circumflex over ( )}

12	IPI:IPI00225062.2	RSSSELSPEVVEK	1629	Srrm2	Isoform 3 of Serine/arginine	1
					repetitive matrix protein 2

13	IPI:IPI00122594.4	TTPLASPSLSPGR{circumflex over ( )}	1630	Ahctf1	AT-hook-containing transcription	1
					factor 1

14	IPI:IPI00136107.1	THSTSSS*IGSGESPFSR{circumflex over ( )}	1631	Ndrg3	Protein NDRG3	1

15	IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	1632	Ndrg3	Protein NDRG3	1

16	IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	1633	Ndrg3	Protein NDRG3	1

17	IPI:IPI00136107.1	THS*TSSSIGSGESPFSR{circumflex over ( )}	1634	Ndrg3	Protein NDRG3	1

18	IPI:IPI00136107.1	THSTSSS*IGSGESPFSR{circumflex over ( )}	1635	Ndrg3	Protein NDRG3	1

19	IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	1636	Ndrg3	Protein NDRG3	1

20	IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	1637	Ndrg3	Protein NDRG3	1

21	IPI:IPI00136107.1	THSTSSSIGSGESPFSR{circumflex over ( )}	1638	Ndrg3	Protein NDRG3	1

22	IPI:IPI00136107.1	THST*SSSIGSGESPFSR{circumflex over ( )}	1639	Ndrg3	Protein NDRG3	1

23	IPI:IPI00551454.3	ER{circumflex over ( )}QES*ESEQELVNK@	1640	Pdcd11	Protein RRP5 homolog	1

24	IPI:IPI00117229.3	TS*PAGGTWSSVVSGVPR	1641	Atxn2	ataxin	2	1

25	IPI:IPI00348442.1	SK@FDS*DEEDEDAENLEA	1642	Sfrs18	splicing factor arginine/serine-rich	1
		VSSGK@			18

26	IPI:IPI00556837.1	DTVIIVSEPSEDEESHDLP	1643	Smarcad1	Isoform 1 of SWI/SNF-related	1
		SVTR			matrix-associated actin-
					dependent regulator of chromatin
					subfamily A containing DEAD/H
					box
1

27	IPI:IPI00153986.2	GTSRPGTPSAEAASTSST	1644	Gtf2f1	General transcription factor IIF	1
		LR			subunit 1

28	IPI:IPI00225062.2	GCSPPKSPEKPPQSTSS*	1645	Srrm2	Isoform	3 of Serine/arginine	1
		ESCPPSPQPTK			repetitive matrix protein 2

29	IPI:IPI00128904.1	VMTIPYQPMPASS*PVICAG	1646	Pcbp1	Poly(rC)-binding protein 1	1
		GQDR

30	IPI:IPI00121251.7	LSTTPSPTNSLHEDGVDD	1647	Tox4	TOX high mobility group box	1
		FRR			family member	4

31	IPI:IPI00320594.5	SQPHSSTSNQETSDSEM	1648	Ranbp10	Ran-binding protein 10	1
		EMEAEHYPNGVLESVSTR{circumflex over ( )}

32	IPI:IPI00808277.2	SAPAS*PNHAGVLSAHSSG	1649	Foxk2	Isoform 1 of Forkhead box	1
		AQTPESLSR{circumflex over ( )}			protein K2

33	IPI:IPI00137166.1	SR{circumflex over ( )}DATPPVSPINMEDQE	1650	Junb	Transcription factor jun-B	1
		R{circumflex over ( )}

34	IPI:IPI00454104.1	TTEAPCSPGSQQPPSPD	1651	Scrib	Isoform 1 of Protein LAP4	1
		ELPANVK@

35	IPI:IPI00225062.2	GCSPPK@SPEK@PPQST	1652	Srrm2	Isoform	3 of Serine/arginine	1
		SSESCPPS*PQPTK@			repetitive matrix protein 2

36	IPI:IPI00153986.2	GTSR{circumflex over ( )}PGTPSAEAASTSS	1653	Gtf2f1	General transcription factor IIF	1
		TLR{circumflex over ( )}			subunit 1

37	IPI:IPI00753321.2	SVSETSEDK@K@DEESD	1654	Bod1l	biorientation of chromosomes in	1
		EEEEEEEEEEPLGATTR{circumflex over ( )}			cell division 1-like

38	IPI:IPI00153986.2	GTSR{circumflex over ( )}PGTPSAEAASTSS	1655	Gtf2f1	General transcription factor IIF	1
		TLR{circumflex over ( )}			subunit 1

39	IPI:IPI00153986.2	GTSR{circumflex over ( )}PGTPSAEAASTSS	1656	Gtf2f1	General transcription factor IIF	1
		TLR{circumflex over ( )}			subunit 1

40	IPI:IPI00929786.1	TASISSSPSEGTPAVGSY	1657	Larp1	Isoform	1 of La-related protein 1	1
		GCT*PQSLPK@

41	IPI:IPI00929786.1	TASISSSPSEGTPAVGSY	1658	Larp1	Isoform	1 of La-related protein 1	1
		GCTPQSLPK@

42	IPI:IPI00130920.1	RSES*PFEGK	1659	Mtap1b	Microtubule-associated protein	1
					1B

43	IPI:IPI00336973.2	GLNLDGTPALSTLGGFS*PA	1660	Ccnl1	Isoform	1 of Cyclin-L1	1
		SK@PS*SPR{circumflex over ( )}

44	IPI:IPI00336973.2	GLNLDGTPALSTLGGFS*PA	1661	Ccnl1	Isoform	1 of Cyclin-L1	1
		S*KPSSPR

45	IPI:IPI00336973.2	GLNLDGTPALSTLGGFSP	1662	Ccnl1	Isoform	1 of Cyclin-L1	1
		ASK@PS*SPR{circumflex over ( )}

46	IPI:IPI00336973.2	GLNLDGTPALSTLGGFSPA	1663	Ccnl1	Isoform	1 of Cyclin-L1	1
		SK@PSSPR{circumflex over ( )}

47	IPI:IPI00379844.4	TAS*EGDGGAAGGAGTAG	1664	Irs2	Insulin receptor substrate 2	1
		GR{circumflex over ( )}PMSVAGSPLSPGPV
		R{circumflex over ( )}

48	IPI:IPI00336973.2	GLNLDGTPALSTLGGFS*PA	1665	Ccnl1	Isoform 1 of Cyclin-L1	1
		SKPSS*PR

49	IPI:IPI00336973.2	GLNLDGTPALSTLGGFSP	1666	Ccnl1	Isoform	1 of Cyclin-L1	1
		ASKPSS*PR

50	IPI:IPI00656285.2	SLVSPIPSPTGTISVPNSC	1667	Foxk1	Forkhead box protein K1	1
		PAS*PR{circumflex over ( )}

51	IPI:IPI00656285.2	SLVSPIPSPTGTISVPNSC	1668	Foxk1	Forkhead box protein K1	1
		PAS*PR

52	IPI:IPI00454104.1	TTEAPCSPGSQQPPSPD	1669	Scrib	Isoform 1 of Protein LAP4	1
		ELPANVK@

53	IPI:IPI00753321.2	SVSETSEDK@K@DEESD	1670	Bod1l	biorientation of chromosomes in	1
		EEEEEEEEEEPLGATTR{circumflex over ( )}			cell division 1-like

54	IPI:IPI00753321.2	SVSETSEDK@K@DEESD	1671	Bod1l	biorientation of chromosomes in	1
		EEEEEEEEEEPLGATTR{circumflex over ( )}			cell division 1-like

55	IPI:IPI00313307.3	SYQNSPSSEDGIR{circumflex over ( )}PLPE	1672	Med1	Isoform	4 of Mediator of RNA	1
		YSTEK@			polymerase II transcription
					subunit
1

56	IPI:IPI00225062.2	GCSPPK@SPEK@PPQS*	1673	Srrm2	Isoform	3 of Serine/arginine	1
		TSSESCPPSPQPTK@			repetitive matrix protein 2

57	IPI:IPI00318938.6	NS*PVAK@TPPK@DLPAIP	1674	Eif4ebp1	Eukaryotic translation initiation	1
		GVTSPTS*DEPPMQASQSQ			factor 4E-binding protein 1
		LPSSPEDK@

58	IPI:IPI00130920.1	SLMSS*PEDLTK@DFEELK	1675	MtaP1b	Microtubule-associated protein	1
		@AEEIDVAK@			1B

59	IPI:IPI00553798.2	SSEVVLSGDDEDYQR{circumflex over ( )}	1676	Ahnak	AHNAK nucleoprotein isoform 1	1

60	IPI:IPI00129264.1	ATSR{circumflex over ( )}PINLGPSS*PNTEIH	1677	Sorbs3	Vinexin	1
		WTPYR{circumflex over ( )}

61	IPI:IPI00129264.1	ATSRPINLGPS*SPNTEIHW	1678	Sorbs3	Vinexin	1
		TPYR

62	IPI:IPI00123410.5	TIS*AQDTLAYATALLNEK@	1679	Usp24	Isoform	1 of Ubiquitin carboxyl-	1
					terminal hydrolase 24

63	IPI:IPI00226441.2	LHYTPPLQSPIT*DGDPLL	1680	Lin9	Isoform 2 of Lin-9 homolog	1
		GQS*PWR{circumflex over ( )}

64	IPI:IPI00309059.7	STSPIIGSPPVR{circumflex over ( )}	1681	Patl1	Protein PAT1 homolog 1	1

65	IPI:IPI00336713.1	CSPVPGLSSSPSGSPLHG	1682	Bcas3	Isoform 1 of Breast carcinoma-	1
		K@			amplified sequence 3 homolog

66	IPI:IPI00320905.7	LGEQGPEPGPTPPQTPT	1683	Arhgap17	Isoform 1 of Rho GTPase-	1
		PPS*TPPLAK			activating protein 17

67	IPI:IPI00309059.7	R{circumflex over ( )}STSPIIGS*PPVR{circumflex over ( )}	1684	Patl1	Protein PAT1 homolog 1	1

68	IPI:IPI00309059.7	RSTSPIIGSPPVR	1685	Patl1	Protein PAT1 homolog 1	1

69	IPI:IPI00309059.7	STSPIIGS*PPVR{circumflex over ( )}	1686	Patl1	Protein PAT1 homolog 1	1

70	IPI:IPI00317599.3	SQEDEEEIST*SPGVSEFVS	1687	Syap1	Synapse-associated protein 1	1
		DAFDTCSLNQEDLRK

71	IPI:IPI00317599.3	SQEDEEEISTS*PGVSEFVS	1688	Syap1	Synapse-associated protein 1	1
		DAFDTCSLNQEDLRK

72	IPI:IPI00656285.2	SLVSPIPSPTGTISVPNS*	1689	Foxk1	Forkhead box protein K1	1
		CPASPR{circumflex over ( )}

73	IPI:IPI00676574.2	RVS*TDLPEGQDVYTAACN	1690	Herc1	hect (homologous to the E6-AP	1
		SVIHR			(UBE3A) carboxyl terminus)
					domain and RCC1 (CHC1)-like
					domain (RLD) 1

74	IPI:IPI00458958.2	DWDK@ES*EGEEPAGGR{circumflex over ( )}	1691	Rrp15	RRP15-like protein	2

75	IPI:IPI00230719.8	R{circumflex over ( )}DS*SDDWEIPDGQITVGQ	1692	Braf	Isoform 1 of B-Raf proto-	2
		R{circumflex over ( )}			oncogene serine/threonine-
					protein kinase

76	IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1693	Eif4ebp1	Eukaryotic translation initiation	2
		GGTLFSTTPGGT*R			factor 4E-binding protein 1

77	IPI:IPI00874995.2	AS*DDLGEPDVFATAPFR	1694	Aak1	Uncharacterized protein	2
					FLJ45252 homolog

78	IPI:IPI00318938.6	VALGDGVQLPPGDYSTT*P	1695	Eif4ebp1	Eukaryotic translation initiation	2
		GGTLFSTT*PGGTR{circumflex over ( )}			factor 4E-binding protein 1

79	IPI:IPI00318938.6	VALGDGVQLPPGDYSTTPG	1696	Eif4ebp1	Eukaryotic translation initiation	2
		GTLFSTTPGGTR			factor 4E-binding protein 1

80	IPI:IPI00318938.6	VALGDGVQLPPGDYS*TTP	1697	Eif4ebp1	Eukaryotic translation initiation	2
		GGTLFSTT*PGGTR{circumflex over ( )}			factor 4E-binding protein 1

81	IPI:IPI00323045.3	S*MDVDLNQAHMEDTPK@	1698	Melk	Maternal embryonic leucine	2
					zipper kinase

82	IPI:IPI00623284.4	IWDPTPSHTPAGAATPGR	1699	Sf3b1	Splicing factor 3B subunit 1	2
		GDTPGHATPGHGGATSS
		AR

83	IPI:IPI00125960.1	TASGSSVTSLEGTR	1700	Ndrg1	Protein NDRG1	2

84	IPI:IPI00318938.6	R{circumflex over ( )}VALGDGVQLPPGDYSTT	1701	Eif4ebp1	Eukaryotic translation initiation	2
		PGGTLFSTTPGGTR{circumflex over ( )}			factor 4E-binding protein 1

85	IPI:IPI00229571.1	ISSK@SPGHMVILNQTK@	1702	Sltm	Isoform	1 of SAFB-like	2
					transcription modulator

86	IPI:IPI00120095.2	DFTK@PQDGDIIAPLIT*PLK@	1703	Samhd1	SAM domain and HD domain-	2
					containing protein 1

87	IPI:IPI00340860.5	TASEGSEAETPEAPK	1704	Larp7	Isoform 1 of La-related protein 7	2

88	IPI:IPI00664670.4	LGS*FGSITR{circumflex over ( )}	1705	Flnc	Isoform	1 of Filamin-C	2

89	IPI:IPI00654192.2	SLS*FSEPQQPPPTVK@	1706	Zfp395	zinc finger protein 395	2

90	IPI:IPI00225062.2	IHTTALTGQS*PPLASGHQG	1707	Srrm2	Isoform	3 of Serine/arginine	2
		EGDAPSVEPGATNIQQPSS			repetitive matrix protein 2
		*PAPSTK

91	IPI:IPI00225062.2	IHTTALTGQS*PPLASGHQG	1708	Srrm2	Isoform 3 of Serine/arginine	2
		EGDAPSVEPGATNIQQPSS			repetitive matrix protein 2
		PAPSTK@

92	IPI:IPI00225062.2	IHTTALTGQS*PPLASGHQG	1709	Srrm2	Isoform	3 of Serine/arginine	2
		EGDAPSVEPGATNIQQPSS			repetitive matrix protein 2
		PAPSTK

93	IPI:IPI00339428.9	MSSHT*ETSSFLQTLTGR	1710	Dock7	Isoform 2 of Dedicator of	2
					cytokinesis protein 7

94	IPI:IPI00225062.2	DGSGTPSRHSLSGSSP	1711	Srrm2	Isoform	3 of Serine/arginine	2
		GMKDTPQT*PSR			repetitive matrix protein 2

95	IPI:IPI00668709.2	DRSSS*APNVHINTIEPVNID	1712	Braf	B-Raf protein	2
		DLIR

96	IPI:IPI00135660.5	RGNNSAVGSNADLT*IEE	1713	Sdpr	Serum deprivation-response	2
		DEEEEPVALQQAQQVR			protein

97	IPI:IPI00135660.5	R{circumflex over ( )}GNNSAVGSNADLTIEE	1714	Sdpr	Serum deprivation-response	2
		DEEEEPVALQQAQQVR{circumflex over ( )}			protein

98	IPI:IPI00404545.2	SLSSPTVTLSAPLEGAK@	1715	Nedd4l	Isoform 3 of E3 ubiquitin-protein	2
					ligase NEDD4-like

99	IPI:IPI00121892.9	R{circumflex over ( )}PPS*PDPNTK@	1716	Spnb2	Isoform	2 of Spectrin beta chain	2
					brain 1

100	IPI:IPI00322707.5	AKQPVIGDQNS*DSDEMLA	1717	Atrx	Transcriptional regulator ATRX	2
		VLK

101	IPI:IPI00929786.1	SLPTTVPESPNYR{circumflex over ( )}	1718	Larp1	Isoform	1 of La-related protein 1	3

102	IPI:IPI00125319.1	TTS*FAESCK@PVQQPSAF	1719	Gsk3b	Glycogen synthase kinase-3 beta	3
		GSMK@

Table 9(b)

	rapa median area
ID	ratio	rapa median SN ratio	KU median area ratio	KU median SN ratio

1	−1.15E+00	−1.14E+00	−0.8183	−0.9318

2	−2.1437	−2.4238	−0.78695	−0.9492

3	−4.758	−4.9643	−0.6283	−0.5475

4	−4.62805	−4.47925	−0.6283	−0.5475

5	−3.6217	−3.885	−0.6283	−0.5475

6	−1.0246	−1.0724	−0.5666	0.1321

7	−1.881	−1.90875	−0.5354	−0.5361

8	−5.2234	−5.13485	−0.4254	−0.6826

9	−1.8021	−2.01515	−0.4151	−0.4336

10	−1.5621	−1.0464	−0.3725	−0.7405

11	−1.4347	−1.3061	−0.3725	−0.7405

12	−1.6324	−1.5779	−0.3224	−0.0637

13	−1.5511	−1.71335	−0.2831	−0.0584

14	−1.78365	−2.00075	−0.2767	0.0092

15	−1.7555	−2.0749	−0.2767	0.0092

16	−1.7499	−1.7737	−0.2767	0.0092

17	−1.5242	−1.76425	−0.2767	0.0092

18	−2.0759	−2.2586	−0.2661	−0.1505

19	−1.9707	−1.7471	−0.2661	−0.1505

20	−1.8874	−1.85315	−0.2661	−0.1505

21	−1.8106	−1.691	−0.2661	−0.1505

22	−1.6838	−1.5207	−0.2661	−0.1505

23	−2.8898	−2.7784	−0.2201	−0.1513

24	−1.0015	−0.8287	−0.2135	−0.3206

25	−3.15205	−2.9693	−0.213	0.1493

26	−1.3868	−0.6688	−0.1992	−0.1577

27	−1.1818	−1.15455	−0.1474	−0.0308

28	−1.7403	−1.8968	−0.1417	0.6197

29	−1.6267	−1.0033	−0.139	−0.2714

30	−1.0844	−1.1514	−0.106	−0.2609

31	−2.78805	−1.56635	−0.1017	−0.8833

32	−3.0877	−2.2749	−0.099	0.6127

33	−2.20425	−2.1077	−0.085	0.023

34	−1.2842	−0.6546	−0.0822	0.1059

35	−1.7868	−2.1396	−0.0816	0.1618

36	−1.2551	−1.22675	−0.0656	−0.106

37	−2.9525	−3.6913	−0.0637	0.7614

38	−1.1888	−1.2446	−0.014	−0.0308

39	−1.2382	−1.25285	−0.0136	0.0107

40	−2.1557	−2.1257	−0.01	−0.0601

41	−4.4981	−3.9942	0.0259	−0.0762

42	−1.7508	−0.9897	0.0386	0.3631

43	−1.896	−2.1685	0.0563	0.2174

44	−1.89265	−2.1434	0.0563	0.1073

45	−1.40465	−1.02655	0.0563	0.2174

46	−1.896	−2.1685	0.0814	0.17005

47	−3.6492	−3.6492	0.0966	0.0223

48	−1.8973	−2.1434	0.1063	0.2306

49	−1.273	−0.7556	0.1063	0.2306

50	−2.8428	−2.9973	0.1159	0.0362

51	−2.8019	−2.9973	0.1159	0.0362

52	−1.2352	−0.0592	0.1368	0.1632

53	−3.106	−3.2482	0.1555	0.3337

54	−2.9525	−3.6913	0.1555	0.3337

55	−1.298	−1.4299	0.2219	0.413

56	−1.7403	−1.8968	0.22985	0.1085

57	−1.3906	−1.2466	0.2704	1.5615

58	−1.1401	−1.2853	0.2835	0.1358

59	−2.4187	−0.9488	0.307	0.366

60	−1.3056	−1.2452	0.3118	0.4053

61	−1.2719	−1.0234	0.3118	0.4053

62	−1.4486	−1.5455	0.3219	0.2377

63	−3.8536	−3.8536	0.3485	0.0243

64	−1.36155	−1.26665	0.3516	0.4392

65	−1.889	−1.727	0.3696	1.0368

66	−2.0572	−2.2717	0.3908	0.1481

67	−1.626	−2.0445	0.51825	0.767

68	−1.29335	−1.2722	0.51825	0.767

69	−1.9492	−2.05475	0.5526	0.865

70	−1.1567	−1.0106	0.5969	0.9976

71	−1.1567	−1.0106	0.5969	0.9976

72	−2.818	−2.9973	1.1365	1.0801

73	−3.4866	−3.4138	1.2714	1.4529

74	0.0401	−0.1138	−3.4595	−5.0415

75	0.011	−0.1502	−3.3715	−3.3715

76	−0.22145	−0.31255	−2.9948	−2.3042

77	0.2389	0.2797	−2.8989	−2.0993

78	−0.2639	−0.32955	−2.8403	−2.8893

79	−0.2514	−0.2836	−2.7755	−2.167

80	−0.2469	−0.3415	−2.5654	−2.1344

81	−0.58015	−0.1451	−2.5295	−2.6811

82	−0.1752	0.3008	−2.3289	−2.6763

83	0.2073	0.2067	−2.0546	−2.0769

84	−0.3079	−0.4553	−1.759	−1.6004

85	0.13985	0.04745	−1.5237	−1.538

86	−0.1736	−0.41725	−1.3706	−0.9742

87	−0.1646	−0.2464	−1.3468	−1.5444

88	0.3745	0.3233	−1.2893	−1.2512

89	−0.5295	−0.8185	−1.1648	−1.0131

90	−0.4199	−0.3301	−1.1227	−0.4758

91	−0.2531	−0.1626	−1.1227	−0.4758

92	−0.1997	−0.1626	−1.1227	−0.4758

93	0.1923	0.3451	−1.1197	−1.2356

94	−0.0084	0.0364	−1.0814	−1.582

95	−0.3065	−0.4384	−1.0477	−1.431

96	−0.18955	−0.4136	−1.0458	−1.0184

97	−0.15495	−0.3246	−1.0458	−1.0184

98	−0.573	−0.2978	−1.0265	−0.7599

99	0.0109	−0.0378	−1.0209	−0.674

100	0.10215	0.0699	−1.011	−2.1889

101	−2.1207	−2.1939	−6.1589	−5.4183

102	−1.71915	−1.62995	−1.07715	−2.06845

Table 9. Classification of mTOR targets identified in the Rapamycin and Ku-0063794 screens. Class 1 represents downstream effectors of rapamycin-sensitive mTORC1. Class 2 represents downstream effectors of rapamycin-insensitive mTORC1 or mTORC2. Class 3 represents the proteins downstream of both mTORC1 and mTORC2. Name of the genes and the sequences of the phosphopeptides are shown. Table 9 is provided in two parts because the colums span more than a single page. Table 9(a) contains colums 1-6 of Table 9 and Table 9(b) contains columns 7-10, wherein corresponding entries are identified in an ID column in each section. Information provided in (a) for ID 1 relates to the information provided in (b) for ID 1 and vice versa, information provided in (a) for ID 2 relates to the information provided in (b) for ID 2 and vice versa, and so on.

TABLE 10

Gene ontology analysis of the hits identified in the rapamycin and Ku-0063794 screens. The Table contains
four subsections: Biological processes that the rapamycin-sensitive and Ku-0063794-sensitive hits overrepresented
are identified in the “Rapa BP GO” and “Ku BP GO” subsection, respectively. Pathways that the Rapamycin-
senstitive and Ku-0063794-sensitive hits overrepresented are shown under “Rapa KEGG pathways” and “Ku
KEGG pathways,” respectively. Subsections are displayed in three parts, (a), (b), and (c), because the Table
columns span more than one page. Corresponding entries are identified by an ID number. For example, entries
under ID number 1 in Rapa BP GO (a), (b), and (c) refer to the same entry, entries under ID number 2 in Rapa BP
GO (a), (b), and (c) refer to the same entry, entries under ID number 1 in Rapa KEGG pathways (a), (b), and (c)
refer to the same entry, and so forth.

Rapa BP GO pathways (a):

ID	Term	Count	%	PValue

1	GO:0010605~negative regulation of macromolecule	20	10.41666667	4.18E−07
	metabolic process
2	GO:0031327~negative regulation of cellular	17	8.854166667	4.17E−06
	biosynthetic process
3	GO:0010558~negative regulation of macromolecule	16	8.333333333	1.31E−05
	biosynthetic process
4	GO:0045934~negative regulation of nucleobase,	15	7.8125	0.0000314
	nucleoside, nucleotide and nucleic acid metabolic
	process

5	GO:0032868~response to insulin stimulus	6	3.125	0.000264
6	GO:0016481~negative regulation of transcription	13	6.770833333	0.000271
7	GO:0016192~vesicle-mediated transport	14	7.291666667	0.000603
8	GO:0032869~cellular response to insulin stimulus	5	2.604166667	0.000783
9	GO:0051253~negative regulation of RNA	11	5.729166667	0.000886
	metabolic process
10	GO:0051248~negative regulation of protein	6	3.125	0.00096
	metabolic process
11	GO:0006898~receptor-mediated endocytosis	4	2.083333333	0.001048574
12	GO:0006468~protein amino acid phosphorylation	16	8.333333333	0.00137311
13	GO:0051276~chromosome organization	12	6.25	0.001936745
14	GO:0016568~chromatin modification	9	4.6875	0.002136023
15	GO:0043434~response to peptide hormone stimulus	6	3.125	0.002310739
16	GO:0032870~cellular response to hormone stimulus	5	2.604166667	0.002406516
17	GO:0008286~insulin receptor signaling pathway	4	2.083333333	0.002457872
18	GO:0045892~negative regulation of transcription,	10	5.208333333	0.003123471
	DNA dependent
19	GO:0006325~chromatin organization	10	5.208333333	0.003622675
20	GO:0046777~protein amino acid autophosphory-	5	2.604166667	0.003851564
	lation
21	GO:0010627~regulation of protein kinase cascade	7	3.645833333	0.004027038
22	GO:0016310~phosphorylation	16	8.333333333	0.004128421
23	GO:0051656~establishment of organelle localization	4	2.083333333	0.004303328
24	GO:0007010~cytoskeleton organization	10	5.208333333	0.004531801
25	GO:0032535~regulation of cellular component size	7	3.645833333	0.004843432
26	GO:0022402~cell cycle process	11	5.729166667	0.00498568
27	GO:0045926~negative regulation of growth	5	2.604166667	0.005003024
28	GO:0032269~negative regulation of cellular protein	5	2.604166667	0.005258125
	metabolic process
29	GO:0065003~macromolecular complex assembly	10	5.208333333	0.005717288
30	GO:0032268~regulation of cellular protein metabolic	9	4.6875	0.006003754
	process
31	GO:0000122~negative regulation of transcription	8	4.166666667	0.007342706
	from RNA polymerase II promoter
32	GO:0032319~regulation of Rho GTPase activity	3	1.5625	0.007895767
33	GO:0046578~regulation of Ras protein signal	7	3.645833333	0.008452332
	transduction
34	GO:0043933~macromolecular complex subunit	10	5.208333333	0.009572997
	organization
35	GO:0006793~phosphorus metabolic process	17	8.854166667	0.009756342
36	GO:0006796~phosphate metabolic process	17	8.854166667	0.009756342
37	GO:0006897~endocytosis	7	3.645833333	0.010081394
38	GO:0010324~membrane invagination	7	3.645833333	0.010081394
39	GO:0006396~RNA processing	11	5.729166667	0.01020022
40	GO:0007169~transmembrane receptor protein	7	3.645833333	0.01110676
	tyrosine kinase signaling pathway
41	GO:0010608~posttranscriptional regulation of gene	6	3.125	0.01469374
	expression
42	GO:0045792~negative regulation of cell size	4	2.083333333	0.014742447
43	GO:0001701~in utero embryonic development	8	4.166666667	0.015417559
44	GO:0051640~organelle localization	4	2.083333333	0.015500935
45	GO:0051493~regulation of cytoskeleton organization	5	2.604166667	0.015757695
46	GO:0007049~cell cycle	13	6.770833333	0.015795779
47	GO:0006417~regulation of translation	5	2.604166667	0.016293879
48	GO:0017148~negative regulation of translation	3	1.5625	0.019054263
49	GO:0034622~cellular macromolecular complex	7	3.645833333	0.019238583
	assembly
50	GO:0008361~regulation of cell size	5	2.604166667	0.020996944
51	GO:0009725~response to hormone stimulus	6	3.125	0.022401918
52	GO:0051056~regulation of small GTPase mediated	7	3.645833333	0.023850905
	signal transduction
53	GO:0030029~actin filament-based process	6	3.125	0.028573614
54	GO:0048729~tissue morphogenesis	7	3.645833333	0.028642761
55	GO:0001932~regulation of protein amino acid	5	2.604166667	0.030261806
	phosphorylation
56	GO:0008104~protein localization	14	7.291666667	0.031319732
57	GO:0000278~mitotic cell cycle	7	3.645833333	0.031803074
58	GO:0034621~cellular macromolecular complex	7	3.645833333	0.032351058
	subunit organization
59	GO:0009719~response to endogenous stimulus	6	3.125	0.03367924
60	GO:0045449~regulation of transcription	31	16.14583333	0.03551581
61	GO:0007163~establishment or maintenance of cell	3	1.5625	0.036247509
	polarity
62	GO:0080135~regulation of cellular response to stress	4	2.083333333	0.036466875
63	GO:0006357~regulation of transcription from RNA	12	6.25	0.037453702
	polymerase II promoter
64	GO:0032314~regulation of Rac GTPase activity	2	1.041666667	0.038299417
65	GO:0040007~growth	6	3.125	0.040063168
66	GO:0022403~cell cycle phase	8	4.166666667	0.040932988
67	GO:0031328-positive regulation of cellular	11	5.729166667	0.042602185
	biosynthetic process
68	GO:0032318~regulation of Ras GTPase activity	4	2.083333333	0.042864282
69	GO:0010604~positive regulation of macromolecule	12	6.25	0.044146336
	metabolic process
70	GO:0022613~ribonucleoprotein complex biogenesis	5	2.604166667	0.044519805
71	GO:0009891~positive regulation of biosynthetic	11	5.729166667	0.044819532
	process
72	GO:0046822~regulation of nucleocytoplasmic	3	1.5625	0.045236118
	transport
73	GO:0006346~methylation-dependent chromatin	2	1.041666667	0.047644564
	silencing
74	GO:0016044~membrane organization	7	3.645833333	0.049524346
75	GO:0007167~enzyme linked receptor protein	7	3.645833333	0.05025049
	signaling pathway
76	GO:0035023~regulation of Rho protein signal	4	2.083333333	0.051230918
	transduction
77	GO:0043009~chordate embryonic development	9	4.6875	0.052874272
78	GO:0006916~anti-apoptosis	4	2.083333333	0.054183284
79	GO:0009792~embryonic development ending in	9	4.6875	0.055294577
	birth or egg hatching
80	GO:0044087~regulation of cellular component	4	2.083333333	0.055689578
	biogenesis
81	GO:0006350~transcription	25	13.02083333	0.055970599
82	GO:0043062~extracellular structure organization	5	2.604166667	0.057303217
83	GO:0043122~regulation of I-kappaB kinase/	3	1.5625	0.057483236
	NF-kappaB cascade
84	GO:0051726~regulation of cell cycle	6	3.125	0.057658627
85	GO:0046907~intracellular transport	9	4.6875	0.059056688
86	GO:0006260~DNA replication	5	2.604166667	0.060776557
87	GO:0048598~embryonic morphogenesis	8	4.166666667	0.060833555
88	GO:0043087~regulation of GTPase activity	4	2.083333333	0.061912428
89	GO:0051129~negative regulation of cellular	4	2.083333333	0.061912428
	component organization
90	GO:0032386~regulation of intracellular transport	3	1.5625	0.062669535
91	GO:0033043~regulation of organelle organization	5	2.604166667	0.063153096
92	GO:0042325~regulation of phosphorylation	7	3.645833333	0.063602017
93	GO:0045786~negative regulation of cell cycle	3	1.5625	0.065320236
94	GO:0001570~vasculogenesis	3	1.5625	0.065320236
95	GO:0045947~negative regulation of translational	2	1.041666667	0.066065335
	initiation
96	GO:0006461~protein complex assembly	6	3.125	0.070461851
97	GO:0070271~protein complex biogenesis	6	3.125	0.070461851
98	GO:0030833~regulation of actin filament	3	1.5625	0.070731878
	polymerization
99	GO:0010557~positive regulation of macromolecule	10	5.208333333	0.073220823
	biosynthetic process
100	GO:0019220~regulation of phosphate metabolic	7	3.645833333	0.073259901
	process
101	GO:0051174~regulation of phosphorus metabolic	7	3.645833333	0.073259901
	process
102	GO:0048589~developmental growth	4	2.083333333	0.073541659
103	GO:0030838~positive regulation of actin filament	2	1.041666667	0.075142677
	polymerization
104	GO:0030308~negative regulation of cell growth	3	1.5625	0.076284196
105	GO:0030036~actin cytoskeleton organization	5	2.604166667	0.077082848
106	GO:0031399~regulation of protein modification	5	2.604166667	0.077082848
	process
107	GO:0000902~cell morphogenesis	7	3.645833333	0.080785789
108	GO:0016477~cell migration	6	3.125	0.08471051
109	GO:0008064~regulation of actin polymerization or	3	1.5625	0.084860387
	depolymerization
110	GO:0030832~regulation of actin filament length	3	1.5625	0.08778162
111	GO:0002009~morphogenesis of an epithelium	5	2.604166667	0.088103636
112	GO:0006333~chromatin assembly or disassembly	4	2.083333333	0.089801855
113	GO:0010638~positive regulation of organelle	3	1.5625	0.090732662
	organization
114	GO:0045941~positive regulation of transcription	9	4.6875	0.091555467
115	GO:0006887~exocytosis	4	2.083333333	0.091694717
116	GO:0008283~cell proliferation	6	3.125	0.092969193
117	GO:0035020~regulation of Rac protein signal	2	1.041666667	0.093035503
	transduction
118	GO:0000226~microtubule cytoskeleton organization	4	2.083333333	0.093604179
119	GO:0060562~epithelial tube morphogenesis	4	2.083333333	0.093604179

Rapa BP GO pathways (b):

ID	Genes

1	BCLAF1,FOXK1,GM5611,ZEB2,SAP30,EIP4EBP1,SORBS3,EIF4EBP2,GM7289,FAM129A,HELLS,
	IBTK,LOC100046628,NACC1,GM9118,TRIM28,RB1,EHMT2,FLNA,FXR1,GM6477,NFIC,CUX1,LOC
	633387,SMARCA4
2	PALM,BCLAF1,NACC1,FOXK1,TRIM28,ZEB2,RB1,EHMT2,FXR1,SAP30,EIF4EBP1,SORBS3,
	EIF4EBP2,CUX1,NFIC,HELLS,SMARCA4
3	BCLAF1,NACC1,FOXK1,TRIM28,ZEB2,RB1,EHMT2,FXR1,SAP30,EIF4EBP2,SORBS3,EIF4EBP2,
	CUX1,NFIC,HELLS,SMARCA4
4	PALM,LOC100046628,BCLAF1,NACC1,FOXK1,GM9118,TRIM28,GM5611,ZEB2,RB1,EHMT2,
	GM6477,SAP30,SORBS3,NPM1,GM7289,CUX1,NFIC,LOC633387,HELLS,SMARCA4
5	ZFP106,IRS2,EIF4EBP1,EIF4EBP2,LOC100048123,LOC544757,MTOR,AKT2
6	NACC1,BCLAF1,FOXK1,TRIM28,ZEB2,RB1,EHMT2,SAP30,SORBS3,NFIC,CUX1,HELLS,SMARCA4
7	MYO5A,GM8786,LOC100048600,SNX17,ARHGAP17,LLGL1,SCRIB,FLNA,DAB2,CTTN,ULK1,RAB1,
	FKBP15,CUX1,ERC1,TRIP10
8	ZFP106,IRS2,EIF4EBP1,EIF4DBP2,LOC100048123,LOC544757,AKT2
9	LOC100046628,FOXK1,GM9118,TRIM28,GM5611,RB1,EHMT2,GM6477,SAP30,SORBS3,NPM1,GM7
	289,CUX1,NFIC,LOC633387,HELLS,SMARCA4
10	IBTK,EIF4EBP1,EIF4EBP2,FAM129A,FLNA,FXR1
11	DAB2,CTTN,GM8786,ULK1,SNX17
12	SRPK2,GM4521,BCR,TRIM28,LOC544757,PKN2,WNK1,RPS6KB1,PRKD2,PTK2,LOC100048123,ULK
	1,AAK1,GSK3B,EEF2K,BMP2K,CDK12,MTOR,AKT2
13	KDM6A,PDS5B,ARID1A,RB1,EHMT2,1600027N09RIK,SMARCC2,LOC675933,CHD1,KDM3B,TINF2,
	HELLS,SMARCA4
14	KDM6A,SMARCC2,CHD1,KDM3B,RB1,1600027N09RIK,EHMT2,HELLS,SMARCA4
15	ZFP106,IRS2,EIF4EBP1,EIF4EBP2,LOC100048123,LOC544757,MTOR,AKT2
16	ZFP106,IRS2,EIF4EBP1,EIF4EBP2,LOC100048123,LOC544757,AKT2
17	ZFP106,IRS2,EIF4EBP1,EIF4EBP2
18	SAP30,SORBS3,FOXK1,TRIM28,RB1,CUX1,EHMT2,NFIC,HELLS,SMARCA4
19	KDM6A,SMARCC2,LOC675933,CHD1,ARID1A,KDM3B,RB1,1600027N09RIK,EHMT2,HELLS,
	SMARCA4
20	PTK2,ULK1,TRIM28,CDK12,MTOR
21	PDCD11,SORBS3,LOC100048123,SQSTM1,LOC544757,ZEB2,RICTOR,FLNA,AKT2
22	SRPK2,GM4521,BCR,TRIM28,LOC544757,PKN2,WNK1,RPS6KB1,PRKD2,LOC100048123,ULK1,
	AAK1,GSK3B,EEF2K,BMP2K,CDK12,MTOR,AKT2
23	MYO5A,LOC100046628,PTK2,GM9118,NPM1,GM5611,GM7289,SCRIB,LOC633387,GM6477
24	INF2,PALM,LOC100046628,FMNL3,GM9118,GM5611,ARHGAP17,RICTOR,INF2Q,FLNA,GM6477,
	PTK2,NPM1,MTAP1B,GM7289,LOC633387,RANBP10
25	LOC100046628,GM9118,FOXK1,GM5611,RICTOR,GM6477,ULK1,NPM1,GM7289,SPNA2,MTOR,
	DNAJC2,LOC633387
26	LOC100046628,PDS5B,ZC3HC1,GM9118,AHCTF1,GM5611,RB1,EHMT2,CD2AP,GM6477,MACF1,
	GSK3B,NPM1,GM7289,DNAJC2,LOC633387,HELLS
27	ATXN2,PTK2,FOXK1,ULK1,DNAJC2
28	IBTK,EIF4EBP1,EIF4EBP2,FAM129A,FXR1
29	SRPK2,LOC100046628,NACC1,GM9118,AHCTF1,GM5611,FLNA,GM6477,EIF3A,PTK2,GTF2F1,
	NPM1,GM7289,LOC633387,HELLS,SMARCA4
30	IBTK,EIF4G1,EIF4EBP1,EIF4EBP2,ZEB2,MTOR,RICTOR,FAM129A,FXR1
31	SAP30,SORBS3,TRIM28,RB1,CUX1,EHMT2,NFIC,SMARCA4
32	MTOR,RICTOR,SCRIB
33	BCR,GBF1,TBC1D4,MTOR,RICTOR,SCRIB,IQSEC1
34	SRPK2,LOC100046628,NACC1,GM9118,AHCTF1,GM5611,FLNA,GM6477,EIF3A,PTK2,GTF2F1,NPM1,
	GM7289,LOC633387,HELLS,SMARCA4
35	SRPK2,GM4521,BCR,TRIM28,LOC544757,PKN2,WNK1,RPS6KB1,PRKD2,PTK2,LOC100048123,UKL1,
	AAK1,GSK3B,CDK12,EEF2K,BMP2K,PPP1R12A,MTOR,AKT2
36	SRPK2,GM4521,BCR,TRIM28,LOC544757,PKN2,WNK1,RPS6KB1,PTK2,LOC100048123,ULK1,
	AAK1,GSK3B,CDK12,EEF2K,BMP2K,PPP1R12A,MTOR,AKT2
37	DAB2,CTTN,GM8786,ULK1,SNX17,FKBP15,TRIP10,SCRIB
38	DAB2,CTTN,GM8786,ULK1,SNX17,FKBP15,TRIP10,SCRIB
39	LOC100046766,SRPK2,PDCD11,LOC100046744,EMG1,SFRS1,LOC100047322,RSL1D1,SFRS8,PCBP1,
	SRRM2,LARP7,LOC100046735,DHX15,SRRM1,LOC100048559
40	ZFP106,GRB10,PTK2,IRS2,EIF4EBP1,EIF4EBP2,FLNA
41	EIF4G1,EIF4EBP1,EIF4EBP2,FAM129A,FLNA,FXR1
42	FOXK1,ULK1,MTOR,DNAJC2
43	MIB1,DAB2,SRFS1,TCFEB,JUNB,LOC100048559,MLL2,MED1,SMARCA4
44	MYO5A,LOC100046628,PTK2,GM9118,NPM1,GM5611,GM7289,SCRIB,LOC633387,GM6477
45	LOC100046628,GM9118,MTAP1B,NPM1,GM5611,SPNA2,GM7289,MTOR,RICTOR,LOC633387,
	GM6477
46	LOC100046628,ZC3HC1,PDS5B,GM9118,LIN9,GM5611,AHCTF1,RB1,EHMT2,CD2AP,GM6477,NUM
	A1,MACF1,GSK3B,NPM1,GM7289,DNAJC2,HELLS,LOC633387
47	EIF4G1,EIF4EBP1,EIF4EBP2,FAM129A,FXR1
48	EIF4EBP1,EIF4EBP2,FXR1
49	SRPK2,PTK2,EIF3A,AHCTF1,HELLS,FLNA,SMARCA4
50	LOC100046628,FOXK1,GM9118,ULK1,NPM1,GM5611,GM7289,MTOR,DNAJC2,LOC633387,GM6477
51	ZFP106,IRS2,EIF4EBP1,EIF4EBP2,LOC100048123,LOC544757,MTOR,AKT2
52	BCR,GBF1,TBC1D4,MTOR,RICTOR,SCRIB,IQSEC1
53	MYO5A,INF2,FMNL3,ARHGAP17,RICTOR,INF2Q,FLNA
54	MIB1,MACF1,TNC,ZEB2,SCRIB,MED1,SMARCA4
55	IBTK,ZEB2,MTOR,RICTOR,FAM129A
56	PALM,LOC100046628,LOC100048600,GM9118,SNX17,GM5611,AHCTF1,FLNA,SCRIB,GM6477,
	POM121,ERBB2IP,MACF1,ULK1,RAB1,GSK3B,NPM1,GM7289,SNX30,ERC1,LOC633387
57	ZC3HC1,PDS5B,AHCTF1,RB1,CD2AP,DNAJC2,HELLS
58	SRPK2,PTK2,EIF3A,AHCTF1,HELLS,FLNA,SMARCA4
59	ZFP106,IRS2,EIF4EBP1,EIF4EBP2,LOC100048123,LOC1000544757,MTOR,AKT2
60	BCLAF1,CARHSP1,FOXK1,FOXK2,AHCTF1,ZEB2,1600027N09RIK,SAP30,SORBS3,GM9791,KDM3B,
	TCFEB,DNAJC2,HELLS,NACC1,PDCD11,TRIM28,CCNL1,RB1,EHMT2,JUNB,FLNA,RFC1,GTF2F1,
	SMARCC2,TMPO,ZFP516,CUX1,NFIC,PBX2,SMARCA4,MED1
61	MACF1,SCRIB,LLGL1
62	LOC100046628,LOC100048123,GM9118,NPM1,LOC544757,GM5611,ZEB2,GM7289,MTOR,LOC633387,
	AKT2,GM6477
63	SAP30,SORBS3,FOXK1,TRIM28,RB1,CUX1,TCFEB,EHMT2,NFIC,PBX2,MED1,SMARCA4
64	MTOR,RICTOR
65	MYO5A,LOC100046628,GM9118,ULK1,NPM1,GM5611,GM7289,EHMT2,LOC633387,MED1,GM6477,
	SMARCA4
66	ZC3HC1,PDS5B,ACHTF1,RB1,EHTMC2,CD2AP,DNAJC2,HELLS
67	LOC100046628,GM9118,FOXK1,TRIM28,AHCTF1,GM5611,RB1,GM6477,GTF2F1,NPM1,GM7289,
	TCFEB,FAM129A,NFIC,PBX2,LOC633387,MED1
68	TBC1D4,MTOR,RICTOR,SCRIB
69	FOXK1,GTF2F1,TRIM28,AHCTF1,MTOR,RB1,RICTOR,TCFEB,FAM129A,NFIC,PBX2,MED1
70	SRPK2,LOC100046628,PDCD11,EIF3A,GM9118,EMG1,NPM1,GM5611,LOC633387,GM6477
71	LOC100046628,GM9118,FOXK1,TRIM28,AHCTF1,GM5611,RB1,GM6477,GTF2F1,NPM1,GM7289,
	TCFEB,FAM129A,NFIC,PBX2,LOC633387,MED1
72	GSK3B,PPP1R12A,FLNA
73	HELLS,SMARCA4
74	DAB2,CTTN,GM8786,ULK1,SNX17,FKBP15,TRIP10,SCRIB
75	ZFP106,GRB10,PTK2,IRS2,EIF4EBP1,EIF4EBP2,FLNA
76	BCR,MTOR,RICTOR,SCRIB
77	MIB1,DAB2,ZEB2,SFRS1,TBFEB,JUNB,LOC100048559,MLL2,MED1,SMARCA4
78	AKT1S1,ZC3HC1,GSK3B,HELLS
79	MIB1,DAB2,ZEB2,SFRS1,TCFEB,JUNB,LOC100048559,MLL2,MED1,SMARCA4
80	PTK2,SPNA2,MTOR,RICTOR
81	BLCAF1,FOXK1,FOXK2,MYEF2,AHCTF1,ZEB2,1600027N09RIK,SAP30,KDM3B,TCFEB,HELLS,
	NACC1,TRIM28,CCNL1,RB1,JUNB,PHF3,RFC1,GTF2F1,SMARCC2,ZFP516,NFIC,CUX1,PBX2,
	MED1
82	MYO5A,PTK2,TNC,LOC100048740,ADAMTS2,SMARCA4
83	PDCD11,SQSTM1,FLNA
84	LOC100046628,BCR,FOXK1,GM9118,NPM1,GM5611,GM7289,RB1,SCRIB,JUNB,LOC633387,
	GM6477
85	MYO5A,LOC100046628,LOC100048600,GM9118,GM5611,FLNA,GM6477,MACF1,ERBB2IP,GSK3B,
	RAB1,NPM1,GM7289,CUX1,ERC1,LOC633387
86	RFC1,LIN9,NFIC,DNAJC2,MED1
87	MIB1,MACF1,ZEB2,EXT1,PBX2,SCRIB,MED1,SMARCA4
88	TBC1D4,MTOR,RICTOR,SCRIB
89	PTK2,ULK1,MTAP1B,SPNA2
90	GSK3B,PPP1R12A,FLNA
91	LOC100046628,GM9118,MTAP1B,NPM1,GM5611,SPNA2,GM7289,MTOR,RICTOR,LOC633387,
	GM6477
92	IBTK,LOC100046628,GM9118,GM5611,ZEB2,RICTOR,RB1,GM6477,NPM1,GM7289,MTOR,
	FAM129A,LOC633387
93	FOXK1,RB1,SCRIB
94	PTK2,JUNB,SMARCA4
95	EIF4EBP1,EIF4EBP2
96	LOC100046628,PTK2,NACC1,GM9118,GTF2F1,NPM1,AHCTF1,GM5611,GM7289,LOC633387,FLNA,
	GM6477
97	LOC100046628,PTK2,NACC1,GM9118,GTF2F1,NPM1,AHCTF1,GM5611,GM7289,LOC633387,FLNA,
	GM6477
98	SPNA2,MTOR,RICTOR
99	FOXK1,GTF2F1,TRIM28,AHCTF1,RB1,TCFEB,FAM129A,NFIC,PBX2,MED1
100	IBTK,LOC100046628,GM9118,GM5611,ZEB2,RICTOR,RB1,GM6477,NPM1,GM7289,MTOR,
	FAM129A,LOC633387
101	IBTK,LOC100046628,GM9118,GM5611,ZEB2,RICTOR,RB1,GM6477,NPM1,GM7289,MTOR,
	FAM129A,LOC633387
102	MYO5A,ULK1,MED1,SMARCA4
103	MTOR,RICTOR
104	FOXK1,ULK1,DNAJC2
105	INF2,FMNL3,ARHGAP17,RICTOR,INF2Q,FLNA
106	IBTK,ZEB2,MTOR,RICTOR,FAM129A
107	DAB2,PTK2,MACF1,ULK1,TRIM28,SCRIB,LLGL1
108	PTK2,TNS1,ULK1,ZEB2,CD2AP,SCRIB
109	SPNA2,MTOR,RICTOR
110	SPNA2,MTOR,RICTOR
111	MIB1,TNC,ZEB2,SCRIB,MED1
112	SMARCC2,CHD1,HELLS,SMARCA4
113	LOC100046628,GM9118,NPM1,GM5611,GM7289,MTOR,RICTOR,LOC633387,GM6477
114	FOXK1,GTF2F1,TRIM28,AHCTF1,RB1,TCFEB,NFIC,PBX2,MED1
115	MYO5A,ARHGAP17,SCRIB,LLGL1
116	IRS2,GSK3B,ZEB2,SCRIB,HELLS,MED1
117	MTOR,RICTOR
118	LOC100046628,PTK2,GM9118,MTAP1B,NPM1,GM5611,GM7289,LOC633387,RANBP10,GM6477
119	MIB1,ZEB2,SCRIB,MED1

Rapa BP GO pathways (c):

	List	Pop	Pop	Fold
ID	Total	Hits	Total	Enrichment	Bonferroni	Benjamini	FDR

1	133	506	13588	4.038158638	0.000424	4.24E−04	6.62E-04
2	133	430	13588	4.039097744	0.00422262	0.002113544	0.006599221
3	133	418	13588	3.910637839	0.013202715	0.003317148	0.020725619
4	133	397	13588	3.860154164	0.031339368	0.006347957	0.049646045
5	133	59	13588	10.38970307	0.234820063	0.037513168	0.4165415
6	133	372	13588	3.57029671	0.240156236	0.033747706	0.427409635
7	133	466	13588	3.069347188	0.458108033	0.065811007	0.950983483
8	133	43	13588	11.87969925	0.548457185	0.069730046	1.232339007
9	133	310	13588	3.625224351	0.593238056	0.072219958	1.393086083
10	133	78	13588	7.858877964	0.622688909	0.072234052	1.508600184
11	133	21	13588	19.46007877	0.655224183	0.073240783	1.647016896
12	133	640	13588	2.554135338	0.752083804	0.08878612	2.151600307
13	133	404	13588	3.034616244	0.860222342	0.115719883	3.022171324
14	133	236	13588	3.896138652	0.885866021	0.119855821	3.328229194
15	133	95	13588	6.452552434	0.904450787	0.122300298	3.595818606
16	133	58	13588	8.807363236	0.913322224	0.120774231	3.742213886
17	133	28	13588	14.59505908	0.917735112	0.117406498	3.820625016
18	133	308	13588	3.317058881	0.958217785	0.140327713	4.831479072
19	133	315	13588	3.243346461	0.974869971	0.154173087	5.583086162
20	133	66	13588	7.739804056	0.980097008	0.156587702	5.925840566
21	133	155	13588	4.613921902	0.983355842	0.15688676	6.187818747
22	133	718	13588	2.276666597	0.984989728	0.154613045	6.33886821
23	133	34	13588	12.01946042	0.987440816	0.154948588	6.59892672
24	133	326	13588	3.133908391	0.990050495	0.156967149	6.937610234
25	133	161	13588	4.441974501	0.992759227	0.161382833	7.397709501
26	133	393	13588	2.859591727	0.993737166	0.16048863	7.607019539
27	133	71	13588	7.194747432	0.993846998	0.156076846	7.632510001
28	133	72	13588	7.094820384	0.995256964	0.158538831	8.006669044
29	133	338	13588	3.022645371	0.997031505	0.166286638	8.676553117
30	133	280	13588	3.283888292	0.997784278	0.169076444	9.092166954
31	133	231	13588	3.53819614	0.999435936	0.197489303	11.01137247
32	133	14	13588	21.89258861	0.999679622	0.205375826	11.79297815
33	133	181	13588	3.95114859	0.99981874	0.212838341	12.57303844
34	133	367	13588	2.783798734	0.999942481	0.231930955	14.12415408
35	133	866	13588	2.005556617	0.999952335	0.230394008	14.37545544
36	133	866	13588	2.005556617	0.999952335	0.230394008	14.37545544
37	133	188	13588	3.804031355	0.999965843	0.231800837	14.81929399
38	133	188	13588	3.804031355	0.999965843	0.231800837	14.81929399
39	133	437	13588	2.571669448	0.999969761	0.229071539	14.98100495
40	133	192	13588	3.724780702	0.99998807	0.241565162	16.20528578
41	133	148	13588	4.141841089	0.999999702	0.300740537	20.88938812
42	133	53	13588	7.710597248	0.999999716	0.295721164	20.95127119
43	133	267	13588	3.061135986	0.999999858	0.301224134	21.80434851
44	133	54	13588	7.56780841	0.99999987	0.296980281	21.90910181
45	133	99	13588	5.15986937	0.9999999	0.295639893	22.230872
46	133	611	13588	2.173732203	0.999999904	0.290960801	22.27849302
47	133	100	13588	5.108270677	0.999999943	0.293467107	22.89881268
48	133	22	13588	13.9316473	0.999999997	0.328676558	26.2532865
49	133	217	13588	3.295658501	0.999999997	0.325881168	26.47233348
50	133	108	13588	4.729880256	1	0.344482216	28.53155823
51	133	165	13588	3.715105947	1	0.357404888	30.13796653
52	133	228	13588	3.136657433	1	0.370168333	31.75922119
53	133	176	13588	3.482911825	1	0.420100589	36.80211781
54	133	238	13588	3.004865104	1	0.41509568	36.87329018
55	133	121	13588	4.221711303	1	0.427055137	38.51844258
56	133	753	13588	1.89948976	1	0.432566345	39.57163653
57	133	244	13588	2.930974978	1	0.431978548	40.04717075
58	133	245	13588	2.919011815	1	0.432065576	40.58206094
59	133	184	13588	3.331480876	1	0.439852811	41.86001448
60	133	2227	13588	1.422149897	1	0.452125635	43.58475519
61	133	31	13588	9.886975503	1	0.453613528	44.25845315
62	133	75	13588	5.448822055	1	0.450365735	44.4589569
63	133	616	13588	1.990235329	1	0.454144887	45.35260328
64	133	4	13588	51.08270677	1	0.456547995	46.10772521
65	133	193	13588	3.176126846	1	0.466771241	47.65120316
66	133	328	13588	2.491839354	1	0.469084762	48.39705082
67	133	552	13588	2.03590498	1	0.477872288	49.80054252
68	133	80	13588	5.108270677	1	0.475050206	50.01764292
69	133	633	13588	1.936785091	1	0.480392471	51.06697676
70	133	137	13588	3.728664728	1	0.478500703	51.36875
71	133	557	13588	2.017629352	1	0.476086095	51.60967499
72	133	35	13588	8.757035446	1	0.474622436	51.94267678
73	133	5	13588	40.86616541	1	0.488078471	53.82618784
74	133	272	13588	2.629256966	1	0.497115601	55.24796589
75	133	273	13588	2.619625988	1	0.497699806	55.7861226
76	133	86	13588	4.751879699	1	0.50004125	56.50311848
77	133	421	13588	2.184058722	1	0.506829853	57.6805401
78	133	88	13588	4.643882433	1	0.511161168	58.59697919
79	133	425	13588	2.163502875	1	0.514070595	59.36036942
80	133	89	13588	4.591703979	1	0.512284313	59.62851936
81	133	1772	13588	1.441385631	1	0.509804241	59.81828305
82	133	149	13588	3.428369582	1	0.51404321	60.70683174
83	133	40	13588	7.662406015	1	0.510980221	60.82544158
84	133	214	13588	2.864450847	1	0.507942029	60.94067925
85	133	431	13588	2.133384505	1	0.512485719	61.84796511
86	133	152	13588	3.360704393	1	0.518823826	62.93700974
87	133	359	13588	2.276666597	1	0.515146769	62.97259844
88	133	93	13588	4.394211335	1	0.51755278	63.64021648
89	133	93	13588	4.394211335	1	0.51755278	63.64021648
90	133	42	13588	7.297529538	1	0.518038348	64.10197022
91	133	154	13588	3.317058881	1	0.516945411	64.39400928
92	133	290	13588	2.466061706	1	0.515677628	64.66313432
93	133	43	13588	7.127819549	1	0.521573895	65.67569163
94	133	43	13588	7.127819549	1	0.521573895	65.67569163
95	133	7	13588	29.19011815	1	0.521941211	66.10627789
96	133	227	13588	2.700407406	1	0.541900011	68.54571328
97	133	227	13588	2.700407406	1	0.541900011	68.54571328
98	133	45	13588	6.811027569	1	0.539576096	68.69004358
99	133	530	13588	1.927649312	1	0.548723845	69.99148633
100	133	301	13588	2.37593985	1	0.545243603	70.01150996
101	133	301	13588	2.37593985	1	0.545243603	70.01150996
102	133	100	13588	4.086616541	1	0.543036347	70.15551118
103	133	8	13588	25.54135338	1	0.547458295	70.9615401
104	133	47	13588	6.521196609	1	0.549517711	71.52373165
105	133	165	13588	3.095921622	1	0.549873189	71.91098296
106	133	165	13588	3.095921622	1	0.549873189	71.91098296
107	133	309	13588	2.314426844	1	0.563991497	73.64287114
108	133	240	13588	2.554135338	1	0.578472896	75.36897392
109	133	50	13588	6.129924812	1	0.575662811	75.43274213
110	133	51	13588	6.009730208	1	0.58511535	76.64517662
111	133	173	13588	2.952757617	1	0.583088756	76.77534105
112	133	109	13588	3.749189487	1	0.586996658	77.45061982
113	133	52	13588	5.894158473	1	0.587580562	77.81289666
114	133	475	13588	1.93576573	1	0.58770496	78.12859078
115	133	110	13588	3.715105947	1	0.584982961	78.1816002
116	133	247	13588	2.481750936	1	0.587000314	78.66120379
117	133	10	13588	20.43308271	1	0.584028864	78.68588514
118	133	111	13588	3.681636524	1	0.583150293	78.89645491
119	133	111	13588	3.681636524	1	0.583150293	78.89645491

Rapa KEGG pathways (a):

ID	Term	Count	%	PValue	minuslog10P

1	mmu04150:mTOR signaling pathway	7	3.51758794	1.57E−06	5.8
2	mmu04012:ErbB signaling pathway	6	3.015075377	3.09E−04	3.51
3	mmu04910:Insulin signaling pathway	7	3.51758794	3.40E−04	3.47
4	mmu05221:Acute myeloid leukemia	4	2.010050251	0.007078848	2.15
5	mmu04510:Focal adhesion	6	3.015075377	0.011465215	1.94
6	mmu04530:Tight junction	5	2.512562814	0.013906132	1.86
7	mmu052GO:Pathways in cancer	7	3.51758794	0.02305239	1.64
8	mmu05215:Prostate cancer	4	2.010050251	0.024273508	1.61
9	mmu03018:RNA degradation	3	1.507537688	0.065096102	1.19
10	mmu05214:Glioma	3	1.507537688	0.072876584	1.14
11	mmu04920:Adipocytokine signaling	3	1.507537688	0.078901015	1.1
	pathway
12	mmu04370:VEGF signaling pathway	3	1.507537688	0.097853719	1.01
13	mmu05220:Chronic myeloid leukemia	3	1.507537688	0.097853719	1.01

Rapa KEGG pathways (b):

ID	Genes

1	IPI00453603,IPI00318938,IPI00399440,IPI00268673,IPI00467843,IPI00221581,IPI00121335
2	IPI00125319,IPI00453603,IPI00318938,IPI00113563,IPI00268673,IPI00121335
3	IPI00125319,IPI00453603,IPI00116923,IPI00318938,IPI00268673,IPI00379844,IPI00121335
4	IPI00453603,IPI00318938,IPI00268673,IPI00121335
5	IPI00403938,IPI00125319,IPI00131138,IPI00113563,IPI00671847,IPI00121335
6	IPI00118143,IPI00380354,IPI00323349,IPI00845596,IPI00121335
7	IPI00125319,IPI00121418,IPI00381495,IPI00113563,IPI00268673,IPI00380817,IPI00121335
8	IPI00125319,IPI00121418,IP100268673,IPI00121335
9	IPI00169888,IPI00309059,IPI00330066
10	IPI00121418,IPI00268673,IPI00121335
11	IPI00268673,IPI00379844,IPI00121335
12	IPI00111169,IPI00113563,IPI00121335
13	IPI00121418,IPI00380817,IPI00121335

Rapa KEGG pathways (c):

ID	List Total	Pop Hits	Pop Total	Fold Enrichment	Bonferroni	Benjamini	FDR

1	41	54	5738	18.14182475	1.10E−04	1.10E−04	0.001624292
2	41	87	5738	9.651808242	0.02142063	0.010768293	0.319622391
3	41	138	5738	7.098974903	0.023507108	0.007897915	0.35107298
4	41	57	5738	9.821138211	0.391818277	0.116903139	7.088295684
5	41	198	5738	4.240946046	0.553895165	0.149082636	11.24945571
6	41	135	5738	5.1833785	0.624786839	0.150728903	13.49142317
7	41	323	5738	3.032998565	0.804570905	0.208022678	21.44454537
8	41	90	5738	6.220054201	0.820953112	0.193469404	22.45479997
9	41	60	5738	6.997560976	0.991011485	0.407576887	50.1731853
10	41	64	5738	6.560213415	0.994992413	0.411206711	54.30152323
11	41	67	5738	6.266472515	0.99682721	0.407268104	57.28300003
12	41	76	5738	5.524390244	0.999259735	0.451577052	65.55273205
13	41	76	5738	5.524390244	0.999259735	0.451577052	65.55273205

Ku BP GO (a):

ID	Term	Count	%	PValue

1	GO:0030032~lamellipodium assembly	3	3.092783505	3.25E−03
2	GO:0006468~protein amino acid phosphorylation	10	10.30927835	7.09E−03
3	GO:0008286~insulin receptor signaling pathway	3	3.092783505	9.83E−03
4	GO:0030029~actin filament-based process	5	5.154639175	0.014807537
5	GO:0006913~nucleocytoplasmic transport	4	4.12371134	0.014880914
6	GO:0051493~regulation of cytoskeleton	4	4.12371134	0.016150894
	organization
7	GO:0006796~phosphate metabolic process	11	11.34020619	0.016942852
8	GO:0006259~DNA metabolic process	7	7.216494845	0.025415897
9	GO:0045947~negative regulation of transla-	2	2.06185567	0.037014038
	tional initiation
10	GO:0007049~cell cycle	8	8.24742268	0.04530749
11	GO:0033043~regulation of organelle organization	4	4.12371134	0.049959502
12	GO:0043484~regulation of RNA splicing	2	2.06185567	0.052460451
13	GO:0010605~negative regulation of macro-	7	7.216494845	0.05420299
	molecule metabolic process
14	GO:0030031~cell projection assembly	3	3.092783505	0.054397233
15	GO:0022402~cell cycle process	6	6.18556701	0.059968997
16	GO:0033554~cellular response to stress	6	6.18556701	0.065899107
17	GO:0045884~regulation of survival gene product	2	2.06185567	0.072676117
	expression
18	GO:0010558~negative regulation of macro-	6	6.18556701	0.073909203
	molecule biosynthetic process
19	GO:0006916~anti-apoptosis	3	3.092783505	0.081129603
20	GO:0031327~negative regulation of cellular	6	6.18556701	0.081184023
	biosynthetic process
21	GO:0046907~intracellular transport	6	6.18556701	0.081807159
22	GO:0009890~negative regulation of biosynthetic	6	6.18556701	0.083692072
	process
23	GO:0006396~RNA processing	6	6.18556701	0.085600178
24	GO:0051129~negative regulation of cellular	3	3.092783505	0.089153114
	component organization
25	GO:0006325~chromatin organization	5	5.154639175	0.08921682
26	GO:0030030~cell projection organization	5	5.154639175	0.092408475
27	GO:0043434~response to peptide hormone	3	3.092783505	0.092426596
	stimulus
28	GO:0016569~covalent chromatin modification	3	3.092783505	0.094076504
29	GO:0043271~negative regulation of ion transport	2	2.06185567	0.097348357
30	GO:0007010~cytoskeleton organization	5	5.154639175	0.098124703

Ku BP GO (b):

ID	Genes

1	CCDC88A,NCK1,VCL
2	ALPK3,BAZ1B,BRAF,MAP3K2,SPNB2,GSK3B,NEK9,RPS6KB1,C230081A13RIK,MELK
3	PHIP,EIF4EBPI,EIF4EBP2
4	EPB4.1L3,NCK1,ARHGEF17,MY09B,FLNC
5	LOCI 00046628,GM9118,SPNB2,GSK3B,NPM1,GM5611,GM7289,MYBBP1A,LOC633387,GM6477
6	LOCI 00046628, CCDC88A,GM9118,SPNB2,MTAP1B,NPM1,GM5611,GM7289,LOC633387,
	GM6477
7	ALPK3,BAZ1B,BRAF,MAP3K2,SPNB2,GSK3B,SYNJ1,NEK9,RPS6KB1,C230081A13R1K,MELK
8	ATRX,TRP53BP1,CCDC88A,MDC1,ORC6L,TOP2B,MYC
9	ElF4EBP1,E1F4EBP2
10	LOC100046628,GM9118,RBL1,GM5611,ZFP318,GM6477,MDC1,GSK3B,NPM1,KIF20B,NEK9,
	GM7289,LOC633387,ERCC6L
11	LOC100046628,CCDC88A,GM9118,SPNB2,MTAP1B,NPM1,GM5611,GM7289,LOC633387,GM6477
12	LOC100046628,GM9118,GPX4,NPM1,GM5611,GM7289,HNRPLL,LOC633387,GM6477
13	LOC100046628,GM9118,RBL1,GM5611,GM6477,EIF4EBP1,EIF4EBP2,NPM1,GM7289,MYC,MYBB
	P1A, LOC633387,SMARCA4
14	CCDC88A,NCK1,VCL
15	LOC100046628,GM9118,GSK3B,NPM1,KIF20B,NEK9,GM5611,GM7289,ZFP318,LOC633387,
	GM6477,ERCC6L
16	ATRX,TRP53BP1,BAZ1B,MDC1,GPX4,GSK3B,HNRPLL
17	AKT1S1,MYC
18	EIF4EBP1,EIF4EBP2,RBL1,MYBBP1A,MYC,SMARCA4
19	AKT1S1,GSK3B,MYC
20	EIF4EBP1,EIF4EBP2,RBL1,MYBBP1A,MYC,SMARCA4
21	LOC100046628,GM9118,SPNB2,GSK3B,NPM1,AKAP12,GM5611,MYO9B,GM7289,MYBBP1A,
	LOC633387,GM6477
22	EIF4EBP1,EIF4EBP2,RBL1,MYBBP1A,MYC,SMARCA4
23	LOC100046766,SF3B1,EXOSC9,LOC100046744,GPX4,LARP7,SRRM2,LOC100046735,POP1,
	LOC675032,LOC100047322,HNRPLL
24	RTN4,SPNB2,MTAP1B
25	BAZ1B,GPX4,RBL1,SETD2,HNRPLL,SMARCA4
26	CCDC88A,NCK1,MTAP1B,TOP2B,VCL
27	PHIP,EIF4EBP1,EIF4EBP2
28	BAZ1B,SETD2,SMARCA4
29	BEST3,NEDD4L
30	EPB4.1L3,LOC100046628,GM9118,NCK1,MTAP1B,NPM1,ARHGEF17,GM5611,GM7289,LOC633387,
	GM6477

Ku BP GO (c):

ID	List Total	Pop Hits	Pop Total	Fold Enrichment	Bonferroni	Benjamini	FDR

1	74	16	13588	34.42905405	0.882872164	8.83E−01	4.76E+3000
2	74	640	13588	2.869087838	0.990722194	0.903678632	10.0912277
3	74	28	13588	19.67374517	0.998498942	0.885501679	13.73744045
4	74	176	13588	5.216523342	0.99994544	0.859597701	19.99753834
5	74	96	13588	7.650900901	0.99994805	0.806833826	20.08660666
6	74	99	13588	7.419055419	0.999977769	0.737959309	21.61357964
7	74	866	13588	2.332376256	0.999986913	0.713301589	22.55196875
8	74	421	13588	3.053091096	0.999999956	0.785616066	31.95670058
9	74	7	13588	52.46332046	1	0.87357656	43.11139373
10	74	611	13588	2.404211085	1	0.869178376	50.01437111
11	74	154	13588	4.769392769	1	0.878480874	53.53582478
12	74	10	13588	36.72432432	1	0.875783402	55.33191067
13	74	506	13588	2.540220062	1	0.869599238	56.5448088
14	74	70	13588	7.869498069	1	0.855869756	56.678091
15	74	393	13588	2.803383536	1	0.869269287	60.34271545
16	74	404	13588	2.727053786	1	0.881875859	63.92395423
17	74	14	13588	26.23166023	1	0.895305874	67.64630011
18	74	418	13588	2.635717057	1	0.888826293	68.28378078
19	74	88	13588	6.25982801	1	0.901699824	71.78744367
20	74	430	13588	2.562162162	1	0.892310062	71.81242243
21	74	431	13588	2.55621747	1	0.884669916	72.09697589
22	74	434	13588	2.538547764	1	0.881167309	72.94148547
23	74	437	13588	2.521120663	1	0.877904651	73.77204657
24	74	93	13588	5.923278117	1	0.879820716	75.2555379
25	74	315	13588	2.914628915	1	0.871223772	75.28140884
26	74	319	13588	2.878081844	1	0.872300131	76.54568607
27	74	95	13588	5.798577525	1	0.863873178	76.55268842
28	74	96	13588	5.738175676	1	0.860545996	77.18216045
29	74	19	13588	19.32859175	1	0.862220501	78.38406819
30	74	326	13588	2.81628254	1	0.856532331	78.66045069

Ku KEGG pathways (a):

ID	Term	Count	%	PValue	minuslogP

1	mmu04012:ErbB signaling pathway	6	6.060606061	2.42E−05	4.62
2	mmu04150:mTOR signaling pathway	4	4.04040404	1.39E−03	2.86
3	mmu05221:Acute myeloid leukemia	4	4.04040404	1.62E−03	2.79
4	mmu04110:Cell cycle	4	4.04040404	0.015586032	1.81
5	mmu04910:Insulin signaling pathway	4	4.04040404	0.01903863	1.72
6	mmu05213:Endometrial cancer	3	3.03030303	0.019575313	1.71
7	mmu04010:MARK signaling pathway	5	5.050505051	0.022772811	1.64
8	mmu03018:RNA degradation	3	3.03030303	0.025605774	1.59
9	mmu04510:Focal adhesion	4	4.04040404	0.048078613	1.32
10	mmu05210:Colorectal cancer	3	3.03030303	0.049516956	1.31
11	mmu04350:TGF-beta signaling	3	3.03030303	0.050554685	1.3
	pathway

Ku KEGG pathways (b):

ID	Genes

1	IPI00453999,IPI00125319,IPI00453603,IPI00230719,IPI00318938,IPI00668709,IPI00131999
2	IPI00453603,IPI00230719,IPI00318938,IPI00668709,IPI00221581
3	IPI00453603,IPI00230719,IPI00318938,IPI00668709,IPI00131999
4	IPI00125319,IPI00124717,IPI00137864,IPI00131999
5	IPI00125319,IPI00453603,IPI00230719,IPI00318938,IPI00668709
6	IPI00125319,IPI00230719,IPI00668709,IPI00131999
7	IPI00230719,IPI00117088,IPI00123474,IPI00668709,IPI00664670,IPI00131999
8	IPI00309059,IPI00119442,IPI00330066
9	IPI00125319,IPI00230719,IPI00405227,IPI00668709,IPI00664670
10	IPI00125319,IPI00230719,IPI00668709,IPI00131999
11	IPI00453603,IPI00137864,IPI00131999

Ku KEGG pathways (c):

ID	List Total	Pop Hits	Pop Total	Fold Enrichment	Bonferroni	Benjamini	FDR

1	25	87	5738	15.82896552	1.38E−03	1.38E−03	0.023962829
2	25	54	5738	17.00148148	0.076050732	0.038777202	1.366086094
3	25	57	5738	16.10666667	0.088400206	0.030380355	1.596618843
4	25	128	5738	7.1725	0.591555799	0.200564953	14.41914204
5	25	138	5738	6.652753623	0.6656829	0.196785171	17.34826426
6	25	52	5738	13.24153846	0.675950335	0.171227632	17.79538958
7	25	265	5738	4.330566038	0.731003969	0.171037042	20.41453368
8	25	60	5738	11.476	0.772029847	0.168744085	22.67212753
9	25	198	5738	4.636767677	0.939707705	0.268063281	38.63940264
10	25	86	5738	8.006511628	0.944686732	0.251342928	39.55225136
11	25	87	5738	7.914482759	0.948025839	0.235718393	40.20324697

TABLE 11

Gene Symbol	Annotation

Rapa replicate 1

1600027N09Rik	RIKEN cDNA 1600027N09 gene
Ahctf1	AT-hook-containing transcription factor 1
Ahnak	AHNAK nucleoprotein isoform 1
Akt1s1	Proline-rich AKT1 substrate 1
Bat2l	Isoform	1 of Protein BAT2-like
Bclaf1	Isoform
2 of Bcl-2-associated transcription factor 1
Bmp2k	Isoform	1 of BMP-2-inducible protein kinase
C130092O11Rik	Isoform 1 of Uncharacterized protein KIAA1680
Carhsp1	Calcium-regulated heat stable protein 1
Ccn/l	Isoform	1 of Cyclin-L1
Chd1	Chromodomain-helicase-DNA-binding protein 1
Cttn	Src substrate cortactin
D6Wsu116e	Isoform
1 of Protein FAM21
Dab2	Isoform p93 of Disabled homolog 2
Dock11	Dedicator of cytokinesis protein 11
Edc4	Isoform	1 of Enhancer of mRNA-decapping protein 4
Eif3a	Eukaryotic translation initiation factor 3 subunit A
Eif4ebp1	Eukaryotic translation initiation factor 4E-binding protein 1
Eif4ebp2	Eukaryotic translation initiation factor 4E-binding protein 2
Eif4g1	Isoform 1 of Eukaryotic translation initiation factor 4 gamma 1
Eif5; LOC100047658	Eukaryotic translation initiation factor 5
Epyc	Epiphycan
Fkbp15	Isoform B of FK506-binding protein 15
Foxk1	Forkhead box protein K1
Gm13697	Novel protein containing MIF4G and MA3 domains
Grb10	Isoform
3 of Growth factor receptor-bound protein 10
Gtf2f1	General transcription factor IIF subunit 1
Hdgfrp2	Isoform	3 of Hepatoma-derived growth factor-related protein 2
Hisppd1	140 kDa protein
Ibtk	Isoform
2 of Inhibitor of Bruton tyrosine kinase
Inf2	Isoform
1 of Inverted formin-2
Iqsec1	IQ motif and Sec7 domain 1 isoform b
Irs2	Insulin receptor substrate 2
Iws1	Isoform	1 of Protein IWS1 homolog
Kdm3b	Isoform
2 of Lysine-specific demethylase 3B
Larp1	Isoform
1 of La-related protein 1
Larp4	Putative uncharacterized protein
Ldhd	22 kDa protein
Luc7l2	Isoform
1 of Putative RNA-binding protein Luc7-like 2
Macf1	Isoform	3 of Microtubule-actin cross-linking factor 1
Mett10d	Isoform 1 of Putative methyltransferase METT10D
Mtap1b	Microtubule-associated protein 1B
Myef2	Isoform	2 of Myelin expression factor 2
Myo5a	Myosin-Va
Ndrg3	Protein NDRG3
Nfic	Isoform	1 of Nuclear factor 1 C-type
Npm1	Nucleophosmin
Numa1	Nuclear mitotic apparatus protein 1
Patl1	Protein PAT1 homolog 1
Pcbp2	Isoform	1 of Poly(rC)-binding protein 2
Pds5b	Isoform	1 of Sister chromatid cohesion protein PDS5 homolog B
Peg3	Isoform 1 of Paternally-expressed gene 3 protein
Phf3	PHD finger protein 3
Phldb1	Isoform	2 of Pleckstrin homology-like domain family B member 1
Phldb2	Isoform	1 of Pleckstrin homology-like domain family B member 2
Pi4k2a	Phosphatidylinositol 4-kinase type 2-alpha
Pkn2	Isoform
1 of Serine/threonine-protein kinase N2
Pla2g4a	Cytosolic phospholipase A2
Plekhm1	Pleckstrin homology domain-containing family M member 1
Ppp1r12a	MCG122391 isoform CRA_e
Rab1	Ras-related protein Rab-1A
Rps6	29 kDa protein
Rps6kb1	Isoform Alpha I of Ribosomal protein S6 kinase beta-1
Rsl1d1	Putative uncharacterized protein
Sap30	Histone deacetylase complex subunit SAP30
Serbp1	Isoform 1 of Plasminogen activator inhibitor 1 RNA-binding protein
Sfrs8	splicing factor arginine/serine-rich 8
Sgta	Isoform	1 of Small glutamine-rich tetratricopeptide repeat-containing protein alpha
Smarca4	Putative uncharacterized protein
Snx17	Sorting nexin-17
Srpk2	serine/arginine-rich protein-specific kinase 2
Srrm2	Isoform	3 of Serine/arginine repetitive matrix protein 2
Syap1	Synapse-associated protein 1
Tbc1d4	140 kDa protein
Tcfeb	Transcription factor EB
Tjp2	Tight junction protein ZO-2
Tmem106b	Transmembrane protein 106B isoform CRA_b
Tmpo	Isoform Beta of Lamina-associated polypeptide 2 isoforms beta/delta/epsilon/gamma
Trim28	Isoform
1 of Transcription intermediary factor 1-beta
Ubxn7	UBX domain-containing protein 7
Usp24	Isoform	1 of Ubiquitin carboxyl-terminal hydrolase 24
Uvrag	UV radiation resistance associated
Wnk1	Serine/threonine-protein kinase WNK1
Zc3hc1	Isoform
1 of Nuclear-interacting partner of ALK
Zfp516	Zinc finger protein 516

Rapa Replicate 2

Mib1	E3 ubiquitin-protein ligase MIB1
rsp6	29 kDa protein
Srrm2	Isoform
3 of Serine/arginine repetitive matrix protein 2
Gm13099	Novel protein similar to preferentially expressed antigen in melanoma-like family
Rnf19b	IBR domain containing 3
Rps6	29 kDa protein
Carhsp1	Calcium-regulated heat stable protein 1
Pgrmc2	Membrane-associated progesterone receptor component 2
FLNA	Filamin-A
Ext1	Exostosin-1
Gm14085	Novel protein similar to solute carrier family 28 (Sodium-coupled nucleoside
	transporter) member 2
Akt1s1	Proline-rich AKT1 substrate 1
Gm9757	Putative uncharacterized protein
Eif4ebp2	Eukaryotic translation initiation factor 4E-binding protein 2
Gon4l	RIKEN cDNA 5830417l10 gene
Mogat1	2-acylglycerol O-acyltransferase 1
Eef2k	Elongation factor	2 kinase
Dhx15	Putative pre-mRNA-splicing factor ATP-dependent RNA helicase DHX15
Srrm1	Isoform
2 of Serine/arginine repetitive matrix protein 1
Setd1a	SET domain containing 1A
D830015G02Rik	Putative uncharacterized protein
Vwa5b1	von Willebrand factor A domain-containing protein 5B1
Ibtk	Isoform
2 of Inhibitor of Bruton tyrosine kinase
Erc1; LOC100048600	Isoform 1 of ELKS/RAB6-interacting/CAST family member 1
Adamts2	A disintegrin and metalloproteinase with thrombospondin motifs 2
Grb10	Isoform	3 of Growth factor receptor-bound protein 10
Foxk1	Forkhead box protein K1
Gm6988	similar to hCG1640785
Tnc	Isoform
1 of Tenascin
Larp1	Isoform	1 of La-related protein 1
Alkbh6	24 kDa protein
Larp7	Isoform
1 of La-related protein 7
Ahnak	AHNAK nucleoprotein isoform 1
Qsox2	Isoform	3 of Sulfhydryl oxidase 2
Wnk1	Serine/threonine-protein kinase WNK1
Megf11	Isoform 4 of Multiple epidermal growth factor-like domains 11
Ahctf1	AT-hook-containing transcription factor 1
Fxr1	Isoform E of Fragile X mental retardation syndrome-related protein 1
Ddx21	Nucleolar RNA helicase 2
Lin9	Isoform	2 of Lin-9 homolog
D6Wsu116e	Isoform
1 of Protein FAM21
Sfrs8	splicing factor arginine/serine-rich 8
Ppfibp2	Isoform	4 of Liprin-beta-2
Hdgfrp2	Isoform	3 of Hepatoma-derived growth factor-related protein 2
Irs2	Insulin receptor substrate 2
Tinf2	Putative uncharacterized protein
Hisppd1	140 kDa protein
Edc3	Enhancer of mRNA-decapping protein 3
Aim1l	Absent in melanoma 1-like
Herc1	hect (homologous to the E6-AP (UBE3A) carboxyl terminus) domain and RCC1
	(CHC1)-like domain (RLD) 1
Fbxw9	F-box and WD-40 domain protein 9
Sfrs18	splicing factor arginine/serine-rich 18
Dnajc2	DnaJ homolog subfamily C member 2
Cux1	cut-like homeobox 1 isoform a
Ranbp10	Ran-binding protein 10
Foxk2	Isoform	1 of Forkhead box protein K2
Bod1l	biorientation of chromosomes in cell division 1-like
Zeb2	Zinc finger E-box-binding homeobox 2
Bcr	Breakpoint cluster region protein
Pdcd11	Protein RRP5 homolog
Hn1l	Hematological and neurological expressed 1-like protein
Palm	Isoform
1 of Paralemmin
Phf3	PHD finger protein 3
Ptk2	Isoform	1 of Focal adhesion kinase 1
1110013L07Rik	Putative uncharacterized protein
Tcfeb	Transcription factor EB
Ehmt2	Isoform	1 of Histone-lysine N-methyltransferase H3 lysine-9 specific 3
Zc3h4	Isoform	2 of Zinc finger CCCH domain-containing protein 4
Ktn1	Isoform	1 of Kinectin
Pbx2	Pre-B-cell leukemia transcription factor 2
Phldb1	Isoform	2 of Pleckstrin homology-like domain family B member 1
Cdgap	Cdc42 GTPase-activating protein
Eif3a	Eukaryotic translation initiation factor 3 subunit A
Tbc1d4	140 kDa protein
Dennd4a	hypothetical protein LOC102442
Wdr91	WD repeat-containing protein 91
Junb	Transcription factor jun-B
Srpk2	serine/arginine-rich protein-specific kinase 2
Eif4b	Eukaryotic translation initiation factor 4B
Gbf1	Golgi-specific brefeldin A-resistance factor 1
Cd2ap	CD2-associated protein
LOC100048559; Sfrs1	Isoform	1 of Splicing factor arginine/serine-rich 1
Ccdc6	coiled-coil domain containing 6
Ndrg3	Protein NDRG3
Ulk1	Putative uncharacterized protein
Arhgap17	Isoform 1 of Rho GTPase-activating protein 17
Spna2	Isoform	2 of Spectrin alpha chain brain
Patl1	Protein PAT1 homolog 1
Bcas3	Isoform	1 of Breast carcinoma-amplified sequence 3 homolog
Atg2a	Autophagy-related protein 2 homolog A
Ccnl1	Isoform
1 of Cyclin-L1
Znrf2	E3 ubiquitin-protein ligase ZNRF2
Larp4	Putative uncharacterized protein
Mtor	Isoform
1 of FKBP12-rapamycin complex-associated protein
Gsk3b	Glycogen synthase kinase-3 beta
Ranbp9	RAN binding protein 9
Kdm6a	Isoform	1 of Lysine-specific demethylase 6A
C130092O11Rik	Isoform 1 of Uncharacterized protein KIAA1680
Mtap1b	Microtubule-associated protein 1B
Prkd2	Serine/threonine-protein kinase D2
Usp36	Ubiquitin specific peptidase 36
Slc4a1ap	solute carrier family 4 (anion exchanger) member 1 adaptor protein
Llgl1	lethal giant larvae homolog 1 isoform 1
Cabin1	calcineurin binding protein 1
Pcbp1	Poly(rC)-binding protein 1
Eif4ebp1	Eukaryotic translation initiation factor 4E-binding protein 1
Rfc1	Rfc1 protein
Pds5b	Isoform
1 of Sister chromatid cohesion protein PDS5 homolog B
St5	Isoform
1 of Suppression of tumorigenicity 5
Eif4g1	Isoform	1 of Eukaryotic translation initiation factor 4 gamma 1
Pom121	Nuclear envelope pore membrane protein POM 121
Smarcc2	Isoform 2 of SWI/SNF complex subunit SMARCC2
Eps8l2	Isoform 1 of Epidermal growth factor receptor kinase substrate 8-like protein 2
Plekhm1	Pleckstrin homology domain-containing family M member 1
B230208H17Rik	Putative GTP-binding protein Parf
Mll2	similar to myeloid/lymphoid or mixed-lineage leukemia 2
Snx30	Sorting nexin-30
Rps6kb1	Isoform Alpha I of Ribosomal protein S6 kinase beta-1
Usp24	Isoform	1 of Ubiquitin carboxyl-terminal hydrolase 24
Micall1	Isoform	1 of MICAL-like protein 1
Med1	Isoform 4 of Mediator of RNA polymerase II transcription subunit 1
Sorbs3	Vinexin
Phldb2	Isoform
1 of Pleckstrin homology-like domain family B member 2
Smarcad1	Isoform 1 of SWI/SNF-related matrix-associated actin-dependent regulator of
	chromatin subfamily A containing DEAD/H box 1
Crkrs	Isoform	2 of Cell division cycle 2-related protein kinase 7
Gtf2f1	General transcription factor IIF subunit 1
Scrib	Isoform	1 of Protein LAP4
2610110G12Rik	Isoform
1 of UPF0635 protein C6orf134 homolog
Mllt4	Isoform
3 of Afadin
Nacc1	Nucleus accumbens-associated protein 1
Zfp106	Isoform 1 of Zinc finger protein 106
Rictor	Isoform	1 of Rapamycin-insensitive companion of mTOR
Tns1	tensin
1
Cttn	Src substrate cortactin
Sbno1	Isoform 2 of Protein strawberry notch homolog 1
Fmnl3	Isoform	1 of Formin-like protein 3
Syap1	Synapse-associated protein 1
Strn3	Striatin-3
Rb1	Retinoblastoma-associated protein
Ehbp1	Isoform
2 of EH domain-binding protein 1
Sqstm1	Isoform	1 of Sequestosome-1
Zc3hc1	Isoform	1 of Nuclear-interacting partner of ALK
Tox4	TOX high mobility group box family member 4
Emg1	Probable ribosome biogenesis protein NEP1
Pcm1	Isoform	1 of Pericentriolar material 1 protein
Trip10	Isoform 3 of Cdc42-interacting protein 4
LOC100048123; Akt2	RAC-beta serine/threonine-protein kinase
Aak1	Isoform 2 of AP2-associated protein kinase 1
Srrm2	Isoform	3 of Serine/arginine repetitive matrix protein 2
Phldb2	Isoform	1 of Pleckstrin homology-like domain family B member 2
Phldb1	Isoform	2 of Pleckstrin homology-like domain family B member 1
Ahctf1	AT-hook-containing transcription factor 1
Ahctf1	AT-hook-containing transcription factor 1
Ahctf1	AT-hook-containing transcription factor 1
Arhgap17	Isoform 1 of Rho GTPase-activating protein 17
Rb1	Retinoblastoma-associated protein
Rb1	Retinoblastoma-associated protein
Tox4	TOX high mobility group box family member 4
Tox4	TOX high mobility group box family member 4
Hells	Isoform	1 of Lymphocyte-specific helicase
Hells	Isoform
1 of Lymphocyte-specific helicase
Hells	Isoform
1 of Lymphocyte-specific helicase
Erbb2ip	Isoform
1 of Protein LAP2
Eif4ebp2	Eukaryotic translation initiation factor 4E-binding protein 2
Eif4ebp2	Eukaryotic translation initiation factor 4E-binding protein 2
Pds5b	Isoform	1 of Sister chromatid cohesion protein PDS5 homolog B
Pds5b	Isoform 1 of Sister chromatid cohesion protein PDS5 homolog B
Pds5b	Isoform 1 of Sister chromatid cohesion protein PDS5 homolog B
Fam129a	Protein Niban
Eif4ebp1	Eukaryotic translation initiation factor 4E-binding protein 1
Ahnak	AHNAK nucleoprotein isoform 1
Ahnak	AHNAK nucleoprotein isoform 1
Ahnak	AHNAK nucleoprotein isoform 1
Eif4ebp1	Eukaryotic translation initiation factor 4E-binding protein 1
Arid1a	AT rich interactive domain 1A
Fam129a	Protein Niban
Fam129a	Protein Niban
Atxn2	ataxin 2
Atxn2	ataxin	2
Ranbp10	Ran-binding protein 10
Ranbp10	Ran-binding protein 10
Ranbp10	Ran-binding protein 10
Ranbp10	Ran-binding protein 10
Ranbp10	Ran-binding protein 10
Tox4	TOX high mobility group box family member 4
Tox4	TOX high mobility group box family member 4

Ku

Myo18a	Isoform 4 of Myosin-XVIIIa
Brd2	Isoform	2 of Bromodomain-containing protein 2
6330577E15Rik	Uncharacterized protein C10orf78 homolog
Best3	Bestrophin-3
Larp1	Isoform	1 of La-related protein 1
Fam62c	Isoform 1 of Extended synaptotagmin-3
Mdn1	Midasin homolog
Rtn4	Isoform
2 of Reticulon-4
Pcdh24	Pcdh24 protein
Synj1	similar to mKIAA0910 protein
Zfp318	zinc finger protein 318 isoform 1
Akt1s1	Proline-rich AKT1 substrate 1
Alpk3	myocyte induction differentiation originator
Zc3h14	Isoform
2 of Zinc finger CCCH domain-containing protein 14
Edc4	Isoform	1 of Enhancer of mRNA-decapping protein 4
Srrm2	Isoform	3 of Serine/arginine repetitive matrix protein 2
Kif20b	Isoform	1 of M-phase phosphoprotein 1
1110007A13Rik	UPF0557 protein C10orf119 homolog
Pi4k2a	Phosphatidylinositol 4-kinase type 2-alpha
Top2b	DNA topoisomerase 2-beta
BC021381	Isoform 2 of Uncharacterized protein KIAA1931
Bbx	Isoform 1 of HMG box transcription factor BBX
Vcl	Vinculin
Eif4ebp1	Eukaryotic translation initiation factor 4E-binding protein 1
Rrp15	RRP15-like protein
D10Wsu102e	Uncharacterized protein C12orf45 homolog
Braf	Isoform 1 of B-Raf proto-oncogene serine/threonine-protein kinase
Atg2b	Isoform 1 of Autophagy-related protein 2 homolog B
Npm1	Nucleophosmin
Pcsk5	proprotein convertase subtilisin/kexin type 5
Mtap1b	Microtubule-associated protein 1B
Ercc6l	DNA excision repair protein ERCC-6-like
Serinc1	Serine incorporator 1
Klf3; LOC100046855	Krueppel-like factor 3
Smarca4	Putative uncharacterized protein
Aak1	Uncharacterized protein FLJ45252 homolog
Eif4ebp2	Eukaryotic translation initiation factor 4E-binding protein 2
Ndrg1	Protein NDRG1
Melk	Maternal embryonic leucine zipper kinase
Arhgef17	Isoform 1 of Rho guanine nucleotide exchange factor 17
Grit	Isoform	2 of Rho/Cdc42/Rac GTPase-activating protein RICS
Mdc1	mediator of DNA damage checkpoint 1
Nfkb2	NF-kB2 splice variant 4
Pcm1	Isoform	1 of Pericentriolar material 1 protein
Mybbp1a	Myb-binding protein 1A
Sf3b1	Splicing factor 3B subunit 1
Atrx	Transcriptional regulator ATRX
Ccdc88a	Isoform 2 of Girdin
Baz1b	Isoform 1 of Tyrosine-protein kinase BAZ1B
Nedd4l	Isoform 3 of E3 ubiquitin-protein ligase NEDD4-like
Orc6l	Origin recognition complex subunit 6
Trp53bp1	Transformation related protein 53 binding protein 1
Map3k2	Mitogen-activated protein kinase kinase kinase 2
Hectd2	Hectd2 protein
Usp10	Ubiquitin carboxyl-terminal hydrolase 10
D830031N03Rik	similar to mKIAA0754 protein
Nck1	non-catalytic region of tyrosine kinase adaptor protein 1
Exosc9	Exosome complex exonuclease RRP45
Dap	Death-associated protein 1
Rps6kb1	Isoform Alpha I of Ribosomal protein S6 kinase beta-1
Lmna	Isoform C2 of Lamin-A/C
Sltm	Isoform
1 of SAFB-like transcription modulator
Sh3pxd2a	Isoform
1 of SH3 and PX domain-containing protein 2A
Flnc	Isoform
1 of Filamin-C
Oxr1	Isoform
2 of Oxidation resistance protein 1
Rin2	Isoform	1 of Ras and Rab interactor 2
Nek9	Serine/threonine-protein kinase Nek9
Pebp1	Phosphatidylethanolamine-binding protein 1
Pop1	Processing of 1 ribonuclease P/MRP family
Serhl	Serine hydrolase-like protein
Epb4.1l3	Isoform	1 of Band 4.1-like protein 3
Hnrpll	Isoform	1 of Heterogeneous nuclear ribonucleoprotein L-like
Samhd1	SAM domain and HD domain-containing protein 1
Zfp828	Zinc finger protein 828
Larp7	Isoform	1 of La-related protein 7
Myc	myc proto-oncogene protein
Myo9a	Isoform 2 of Myosin-IXa
Gsk3b	Glycogen synthase kinase-3 beta
Zfp395	zinc finger protein 395
Bend3	BEN domain-containing protein 3
Akap12	Isoform 1 of A-kinase anchor protein 12
Eif4g1	Isoform 1 of Eukaryotic translation initiation factor 4 gamma 1
Eif4b	Eukaryotic translation initiation factor 4B
Dock7	Isoform 2 of Dedicator of cytokinesis protein 7
Patl1	Protein PAT1 homolog 1
Slc7a11	Cystine/glutamate transporter
Myo9b	Isoform 1 of Myosin-IXb
Setd2	SET domain containing 2
Gphn	Gephyrin
Erf	ETS domain-containing transcription factor ERF
Spnb2	Isoform
2 of Spectrin beta chain brain 1
Phip	PH-interacting protein
Sdpr	Serum deprivation-response protein
Tcof1	Treacle protein
Pwp1	Periodic tryptophan protein 1 homolog
Rb/1	Isoform Long of Retinoblastoma-like protein 1
Eef1b2	Elongation factor 1-beta
Phactr4	Isoform
1 of Phosphatase and actin regulator 4
C230081A13Rik	Tyrosine-protein kinase-protein kinase SgK269
Ahnak2	Putative uncharacterized protein

Table 11. Proteins with downregulated phosphorylation identified in the rapamycin and Ku-0063794 screen.

Claims

1. A method for determining mTOR kinase activity in a cancer cell, the method comprising

(a) obtaining a cancer cell from a subject diagnosed to have a cancer,

(b) determining the level of Grb10 phosphorylation in the cell, and

(c) comparing the level of Grb10 phosphorylation to a reference level, wherein if the level of Grb10 phosphorylation in the cell is higher than the reference level, then the cell is determined to exhibit an elevated level of mTOR kinase activity.

2. The method of claim 1, further comprising selecting a method of treatment based on the cell exhibiting an elevated level of mTOR kinase activity.

3. The method of claim 2, wherein if the cell is determined to exhibit an elevated level of mTOR kinase activity, then the selected method of treatment comprises administering an effective amount of an mTOR kinase inhibitor to the subject.

4. The method of claim 3, wherein the method of treatment further comprises administering an effective amount of a compound that stabilizes Grb10 or that inhibits the degradation of Grb10 and/or an effective amount of a PI3K inhibitor to the subject.

5. The method of claim 3, wherein the mTOR inhibitor is an allosteric mTOR kinase inhibitor or a catalytic mTOR kinase inhibitor.

6. The method of claim 5, wherein the allosteric mTOR kinase inhibitor is rapamycin or a rapamycin analog, or wherein the catalytic mTOR kinase inhibitor is an ATP-competitive mTOR kinase inhibitor.

7. The method of claim 3, wherein the mTOR inhibitor is an mTORC1 inhibitor or an mTORC1/2 inhibitor.

8-10. (canceled)

11. The method of claim 3, wherein the mTOR inhibitor is a dual PI3K/mTOR kinase inhibitor.

12-13. (canceled)

14. The method of claim 4, wherein the PI3K inhibitor is a dual PI3K/mTOR kinase inhibitor.

15. (canceled)

16. The method of claim 3, wherein the method of treatment further comprises administering an effective amount of an Akt inhibitor to the subject.

17. (canceled)

18. The method of claim 4, wherein the compound that inhibits the degradation of Grb10 is a ubiquitin ligase inhibitor.

19. The method of claim 18, wherein the ubiquitin ligase inhibitor is an E3 ubiquitin ligase inhibitor.

20. The method of claim 2, further comprising carrying out the selected method of treatment.

21. A method for selecting a treatment of a cancer, the method comprising,

(a) obtaining a cancer cell from a subject diagnosed to have a cancer exhibiting an elevated level of mTOR activity,

(b) determining the level of Grb10 expression in the cell, and

(c) comparing the level of Grb10 expression to a reference level, wherein if the level of Grb10 expression in the cell is higher than the reference level, then the cell is determined to exhibit a high risk of expressing an elevated level of PI3K, Akt, and/or MAPK activity upon being contacted with an mTORC1 inhibitor.

22. The method of claim 21, the method further comprising selecting a method of treatment based on the cell exhibiting a high risk of expressing an elevated level of PI3K, Akt, and/or MAPK activity upon being contacted with an mTORC1 inhibitor.

23. The method of claim 22, wherein the method of treatment comprises

administering (i) an effective amount of an mTOR kinase inhibitor and/or (ii) an effective amount of an IGF1R, EGFR, PI3K, Akt, MEK, and/or RSK inhibitors and/or of a compound stabilizing Grb10 to the subject, or

administering an effective amount of a dual or multi-target inhibitor that inhibits mTOR and inhibits IGFR, EGFR, PI3K, Akt, MEK, and/or RSK and/or of a compound stabilizing Grb10 to the subject.

24-40. (canceled)

41. A method for determining mTOR kinase activity in a cell, the method comprising

(a) determining the level of phosphorylation of a plurality of phosphorylation sites disclosed in Table 1, 2, 3, 7, or 8, or 11 in the cell; and

(b) comparing the level of phosphorylation determined in step (a) to a reference level, wherein

(i) if the level of phosphorylation determined in step (b) is higher than the reference level, then the cell is determined to exhibit an elevated level of mTOR kinase activity, or

(ii) if the level of phosphorylation determined in step (b) is equal or lower than the reference level, then the cell is determined to not exhibit an elevated level of mTOR kinase activity.

42-78. (canceled)

79. A method for determining mTOR kinase activity in a cell, the method comprising

(a) determining the level of phosphorylation of a phosphorylation site disclosed in Table 1, 2, 3, 7, 8, or 11 in the cell; and

(b) comparing the level of phosphorylation determined in step (a) to a reference level, and

80-126. (canceled)

127. A method of identifying an mTOR kinase inhibitor, the method comprising

(a) contacting an mTOR kinase with a polypeptide comprising a phosphorylation site disclosed in Table 1, 2, 3, 7, 8, or 11 under conditions suitable for the mTOR kinase to phosphorylate the phosphorylation site in the presence of a candidate agent,

(b) determining the level of phosphorylation at the phosphorylation site,

(c) comparing the level of phosphorylation determined in step (b) to a reference level, wherein if the level determined in step (b) is lower than the reference level, then the candidate agent is identified as an mTOR kinase inhibitor.

128-139. (canceled)

140. A method for determining mTOR kinase activity in a cell, the method comprising

(a) obtaining a cell from a subject,

(b) determining the level of phosphorylation of a protein selected from the group consisting of Grb10, FOXK1, ZEB2, NDRG3, LARP1, SRPK2, CDK12, MIB1, and IBTK in the cell, and

(c) comparing the level of phosphorylation of the protein to a reference level, wherein if the level of phosphorylation of the protein in the cell is higher than the reference level, then the cell is determined to exhibit an elevated level of mTOR kinase activity.

141-148. (canceled)